CN1977886A

CN1977886A - Medicinal composition of oxymatrine, ganoderma lucidum and astragalus

Info

Publication number: CN1977886A
Application number: CN 200610163454
Authority: CN
Inventors: 黄振华
Original assignee: Individual
Current assignee: Haian Su Fu Technology Transfer Center Co Ltd
Priority date: 2005-12-05
Filing date: 2006-12-04
Publication date: 2007-06-13
Anticipated expiration: 2026-12-04
Also published as: CN1977886B

Abstract

The present invention discloses a new medicine composition and its preparation method. Said medicine composition is formed from (by weight portion) 100-1600 portions of kurarinone, 500-24000 portions of ganoderma and 1000-80000 portions of astragalus root. Besides, it also can be made up by using 100-1600 portions of kurarinone, 2.5-700 portions of ganoderma polysaccharide and 5-1600 portions of astragalus polysaccharide. Said medicine composition can be made into various dosage forms, can be used for preparing the medicines capable or curing acute and chronic hepatitides and active hepatocirrhosis, etc.

Description

A kind of pharmaceutical composition of making by kurarinone, Ganoderma and the Radix Astragali

[technical field]

The invention belongs to medical technical field, be specifically related to a kind of pharmaceutical composition of mainly making by kurarinone, Ganoderma or its extract and the Radix Astragali or its extract, and its production and use.

[background technology]

Hepatitis is a kind of commonly encountered diseases and frequently-occurring disease, and according to incompletely statistics, China's hepatitis and virus carrier account for 10% of total population.Wherein based on viral hepatitis, viral hepatitis can be divided into again: acute hepatitis, chronic hepatitis, hepatitis gravis and gallbladder type hepatitis.Chronic hepatitis can be divided into chronic persistent hepatitis and active hepatitis again.Hepatitis is a kind of infectious disease, because it is popular extensively, hazardness is big, sickness rate is high, is one of difficult and complicated illness of generally acknowledging both at home and abroad at present, with the hepatitis B is example, cure rate is 10% only, and the overwhelming majority transfers chronic hepatitis to, serious hepatic ascites, the hepatocarcinoma of being converted into.Wherein the formation and development of hepatic fibrosis is to influence one of crucial pathological change of prognosis and state of an illness commentaries on classics danger, also is the most complicated knotty problem in the clinical chronic hepatopathy treatment.At present the therapy of treatment chronic hepatitis B mainly contains: antiviral drugs therapy, immunoregulation pharmacotherapy, improve liver microcirculation therapy, protective therapy, differentiation of tcm etc.Though it is many that domestic and international medicine for the treatment of hepatitis in recent years has, most very not generally acknowledged curative effect, and curative effect is unsatisfactory, treat the back state of an illness in addition easily repeatedly, and price is very expensive.So far, still lack the anti-hepatitis determined curative effect less medicine of side effect again on the market.

Kurarinone is the alkaloid that extracts from leguminous plant Herba Sophorae alopecuroidis, Radix Sophorae Flavescentis, and Main Ingredients and Appearance is the N-oxysophocarpine of oxymatrine and minute quantity.Kurarinone has recorded into the 16th 363 pages of national drug standards chemical drugs provincial standard rising national standards of National Drug Administration (Chinese Pharmacopoeia Commission's volume), and wherein regulation contains oxymatrine (C ₁₅H ₂₄N ₂O ₂) must not be less than 98.0%.Kurarinone is mainly used in the leukopenia that low leukocyte counts that the treatment of chronic viral hepatitis B and tumor radiotherapy, chemotherapy cause and other reasons cause.Kurarinone has good antihepatitic activity: use the clinical symptoms of Oxymatrine in Treating Chronic Hepatitis B, as weak, poor appetite and abdominal distention etc., effective percentage is about 90%.The structural formula of oxymatrine is as follows:

Oxymatrine

Ganoderma is Polyporaceae fungus Ganoderma lucidum (Leyss. Ex Fr.) Karst. Ganoderma lucidum (Ley-ss.ex Fr.) Karst. or Ganoderma Ganodermasinense Zhao, the dry sporophore of Xu et Zhang.The property sweet, flat, GUIXIN, lung, liver, kidney channel.Have invigorating QI and tranquilization, the effect of relieving cough and asthma is mainly used in dizzy sleeplessness, shortness of breath and palpitation, cough due to consumptive disease.Ganoderma is widely known Chinese medicine, is commonly called as " Herba mesonae chinensis ".Beginning is stated from Shennong's Herbal, and " tonifying liver QI invigorating " " the hard muscles and bones " that be considered to can " the beneficial motive " " to pacify smart soul " classified as top grade.Compendium of Material Medica thinks that Ganoderma has " strengthening by means of tonics " " life lengthening " " sharp joint " effects such as " controlling deafness ".Ganoderma can strengthening the body resistance, raise immunity, and human body immunity improving power, two-ways regulation function of human body balance is regulated the internal body vigor on the whole, regulates human body metabolism function, improves autoimmunity, promotes internal organs or organ function normalization.The main effective ingredient of Ganoderma is a ganoderan, and ganoderan has the effect of significant enhance immunity.

The Radix Astragali is the dry root of leguminous plant Radix Astagali Astragalus membranaceus (Fisch.) Bge.var.mongholicus (Bge.) Hsiao or Radix Astragali Astragalus membranaceus (Fisch.).Sweet, temperature is returned lung, spleen channel, has the effect of invigorating QI to consolidate the body surface resistance, diuresis detoxification, evacuation of pus, expelling pus and promoting granulation, and it is weak to be mainly used in the deficiency of vital energy, anorexia and loose stool, sinking of QI of middle-JIAO, chronic diarrhea proctoptosis, the metrorrhagia of having blood in stool, exterior deficiency spontaneous perspiration, deficiency of vital energy edema, carbuncle is difficult bursts, and burst for a long time and do not hold back, the blood deficiency dull yellowish colored skin, interior-heat is quenched one's thirst; Chronic nephritis albumen disease, diabetes.The modern pharmacology experiment shows that the Radix Astragali can promote immunologic function, and main effective ingredient is polysaccharide and saponin.Astragalus polysaccharides can significantly improve the immunity of body, strengthens the physiological metabolism effect of human body cell, significantly promotes myeloid element DNA synthetic, accelerates the nucleated cell fission process, and blood sugar level in the body is had dual regulation.Radix Astragali saponin has the effect of anti-liver injury, can alleviate the pathological changes that liver poisoning causes.

Utilize the interaction of kurarinone, Ganoderma or its extract and the Radix Astragali or its extract at present, the application that composition of prescription is used for preparing the medicine for the treatment of hepatic disease yet there are no report.

[summary of the invention]

In order to meet clinical needs, to the invention provides a kind of new being mainly used in and prepare the pharmaceutical composition of the medicine of treatment hepatic disease, and its preparation method is provided.

Pharmaceutical composition of the present invention is mainly made by kurarinone, Ganoderma and the Radix Astragali, and its parts by weight are: 100～1600 parts of kurarinones, 500～24000 parts of Ganodermas, 1000～80000 parts of the Radixs Astragali; Be preferably: 200～800 parts of kurarinones, 1000～12000 parts of Ganodermas, 10000～40000 parts of the Radixs Astragali; The best is: 400 parts of kurarinones, 3000～9000 parts of Ganodermas, 20000 parts of the Radixs Astragali.

Ganoderma in the aforementioned pharmaceutical compositions, the Radix Astragali can be with The suitable solvent respectively or mix through extracting processing and obtain its extract, and total extract is made clinically arbitrary or pharmaceutically acceptable dosage form with kurarinone and mixing acceptable accessories again." extract separately " and be meant that Ganoderma, each flavor medical material of the Radix Astragali extract separately by different technology respectively and obtain extract, each extract are mixed obtaining total extract again." mixed extraction " is meant, Ganoderma, the Radix Astragali two flavor medical materials extract together and obtain total extract.Contained main effective ingredient is polysaccharide or polysaccharide and total saponins in the total extract, and main content of effective is not less than 30% in the total extract.

Ganoderma can obtain ganoderan through extracting processing with The suitable solvent, preferred sour water of solvent wherein or ethanol, the extracting method of ganoderan can be in infusion process, percolation, decocting method, reflux extraction, the continuous extraction one or more, but also reference literature method preparation.

The preferred extracting method that the invention provides Ganoderma is as follows:

Get the Ganoderma medical material, be ground into coarse grain, decoct three times after adding 4 times in water amount moistening 24h, add for the first time 12 times of water gagings and decocted 3 hours, add 10 times of water gagings for the second time and decocted 2 hours, add 10 times of water gagings for the third time and decocted 1 hour, collecting decoction filters, and it is 1.10～1.15 that filtrate is concentrated into relative density, filter, filtrate is removed albumen with the Sevage method, filters, it is 60% to containing the alcohol amount that filtrate adds ethanol, stirs evenly, and cold preservation was left standstill 24 hours, filter, collect filter cake, add an amount of water stirring and dissolving, filter, it is 85% to containing the alcohol amount that filtrate adds ethanol, stirs evenly, cold preservation was left standstill 24 hours, filtered, and collected filter cake, add proper amount of acetone, the washing with alcohol cyclic washing repeatedly, sucking filtration, precipitation adds suitable quantity of water makes dissolving, ultrafiltration, collect molecular weight greater than 50000 daltonian parts, concentrate, vacuum drying, promptly.

The yield of the ganoderan by this prepared is 0.5～3%, and the content of polysaccharide is not less than 80% in the ganoderan.

Except that adopting said method, Ganoderma also can prepare by the following method, but is not limited only to following method:

Method one: get the Ganoderma medical material, be ground into coarse grain, decoct with water three times after adding 3 times in water amount moistening 24h, add for the first time 12 times of water gagings and extracted second 3 hours, add 10 times of water gagings for three times and extracted collecting decoction 2 hours, filter, it is 1.03～1.08 that filtrate is concentrated into relative density, filters, filtrate is removed albumen with the Sevage method, filters, and filtrate adds ethanol makes that to contain the alcohol amount be 70%, stir evenly, cold preservation was placed 24 hours, filtered, collect filter cake, add an amount of water stirring and dissolving, filter, filtrate adds ethanol makes that to contain alcohol amount be 85%, stirs evenly, and cold preservation was placed 24 hours, filter, collect filter cake, add proper amount of acetone, the washing with alcohol cyclic washing repeatedly, sucking filtration, precipitation adds suitable quantity of water makes dissolving, cold preservation was placed 24 hours, filtered to collect filtrate, concentrated, vacuum drying, promptly.

The yield of the ganoderan by this prepared is 2～4%, and the content of ganoderan is not less than 60%.

Method two: get the Ganoderma medical material, be ground into coarse grain, decoct secondary after adding 3 times in water amount moistening 24h, add 12 times of amounts of water for the first time in 3 hours, added 10 times of amounts of water, collecting decoction for the second time in 2 hours, filter, it is 1.02～1.06 that filtrate is concentrated into relative density, filters, filtrate is removed albumen with the Sevage method, filters, and filtrate adds ethanol makes that to contain the alcohol amount be 65%, stir evenly, cold preservation was placed 24 hours, filtered, collect filter cake, add suitable quantity of water and make dissolving, filter, filtrate adds ethanol makes that to contain alcohol amount be 80%, stirs evenly, and cold preservation was placed 24 hours, filter, collect filter cake, add suitable quantity of water and make dissolving, cold preservation was placed 48 hours, filtered, and filtrate concentrates, vacuum drying, promptly.

The yield of the ganoderan by this prepared is 3～5%, and the content of ganoderan is not less than 50%.

Modern pharmacology experiment shows that the astragalus polysaccharides in the Radix Astragali has the immunity of raising activity, and Radix Astragali total saponins has liver-protecting activity, so " extracting separately " method of the Radix Astragali can adopt different extracting method because of the difference of its active component, and is as follows respectively:

Be that the preparation method of the Radix Astragali of main effective ingredient is as follows with polysaccharide in the present composition:

Get Milkvetch Root, add 7 times of amount 70% ethanol extraction secondaries, each 2 hours, extracting solution discards, the slag extracting in water secondary of getting it filled, and extracting solution merges, the ratio that is concentrated into extracting liquid volume and raw medicinal herbs is 1.05: 1, add ethanol precipitation and make and contain the alcohol amount and reach 70%, filter, must precipitate, precipitation 70% washing with alcohol, the dissolving of reuse suitable quantity of water is filtered, and filtrate is crossed macroporous resin, the water eluting, collect water lotion, water lotion is concentrated into medicine liquid volume, add ethanol and make and contain alcohol and measure and reach 70% with the medical material ratio is 1: 2.5, must precipitate, to precipitate with 95% ethanol and washing with acetone dehydration, drying under reduced pressure (60 ℃), promptly.

The yield of the Radix Astragali extract by this prepared is 0.5～2%, and wherein the content of polysaccharide is not less than 50%.

Except that adopting said method, the Radix Astragali also can prepare by the following method, but is not limited only to following method:

Method one: get Milkvetch Root, add the water reflux, extract, three times, each 2 hours, merge extractive liquid, filters, and it is 1.15～1.23 that filtrate decompression is concentrated into relative density, add ethanol and make and contain alcohol amount and reach 80%, left standstill 24 hours, filter, precipitation is dissolved in water, and filters, and adds ethanol again and makes and contain the alcohol amount and reach 85%, left standstill 24 hours, filter, collecting precipitation, vacuum drying are promptly.

The yield of the Radix Astragali extract by this prepared is 2～4%, and wherein the content of polysaccharide is not less than 40%.

Method two: get Milkvetch Root, add 10 times of amount 80% ethanol extractions 4 hours, extracting solution discards, the slag extracting in water secondary of getting it filled, each 2 hours, add 10 times of amounts of water at every turn, extracting solution merges, and filters, filtrate adds ethanol to be made and contains alcohol amount and reach 85%, filters, and filtrate decompression is concentrated into the thick paste shape, and spray drying promptly.

The yield of the Radix Astragali extract by this prepared is 3～4%, and wherein the content of polysaccharide is not less than 30%.

Be that the preparation method of the Radix Astragali of main effective ingredient is as follows with total saponins in the present composition:

Get the Radix Astragali, decoct with water three times, each 1.5 hours, add for the first time 10 times of amounts of water, be 8 times of amounts two, three times, collecting decoction, filter, it is 1.20～1.25 (60 ℃) that filtrate is concentrated into relative density, handles 2 times with ethanol precipitation, containing the alcohol amount in the solution for the first time is 75%, be 85% for the second time, each all cold preservation is placed, and reclaims ethanol and be concentrated into every 1ml to be equivalent to crude drug 10g, be added on the macroporous adsorptive resins of having handled well, earlier with the water of 2 times of volumes towards post, 70% ethanol elution of 4 times of volumes of reuse, collection eluent, concentrating under reduced pressure, vacuum drying, promptly.

The yield of the Radix Astragali extract by this prepared is 0.5～2%, and total saponin content is not less than 50%, and wherein Astragaloside content is not less than 2.0%.

Method one: get the Radix Astragali, decoct with water three times, each 2 hours, collecting decoction, filter, it is 1.20～1.25 (60 ℃) that filtrate is concentrated into relative density, handles 2 times with ethanol precipitation, containing amount of alcohol in the solution for the first time is 60%, be 85% for the second time, each all cold preservation is placed, and reclaims ethanol and be concentrated into every 1ml to be equivalent to crude drug 10g, concentrating under reduced pressure, vacuum drying, promptly.

The yield of the Radix Astragali extract by this prepared is 3～5%, and total saponin content is not less than 30%, and wherein Astragaloside content is not less than 1%.

Method two: get the Radix Astragali, decoct with water three times, each 1.5 hours, collecting decoction filtered, it is 1.20～1.25 (60 ℃) that filtrate is concentrated into relative density, handle to make that to contain amount of alcohol be 60% for 1 time with ethanol precipitation, cold preservation is placed, and reclaims ethanol and be concentrated into every 1ml to be equivalent to crude drug 10g, and concentrating under reduced pressure, vacuum drying, promptly.

The yield of the Radix Astragali extract by this prepared is 2～4%, and total saponin content is for being not less than 40%, and wherein Astragaloside content is not less than 1%.

Pharmaceutical composition of the present invention, available ganoderan, astragalus polysaccharides or Radix Astragali total saponins replace the Ganoderma and the Radix Astragali to feed intake respectively making, (yield of ganoderan is 0.5～3% with respect to the yield of medical material according to extract, the yield of astragalus polysaccharides or total saponins is 0.5～2%) calculate, following two kinds of different proportionings can be arranged, and its parts by weight are respectively:

Proportioning 1: 100～1600 parts of kurarinones, 2.5～700 parts of ganoderans, 5～1600 parts of astragalus polysaccharidess; Be preferably: 200～800 parts of kurarinones, 5～350 parts of ganoderans, 50～800 parts of astragalus polysaccharidess; The best is: 400 parts of kurarinones, 15～250 parts of ganoderans, 100～400 parts of astragalus polysaccharidess.

Proportioning 2: 100～1600 parts of kurarinones, 2.5～700 parts of ganoderans, 5～1600 parts of Radix Astragali total saponinss; Be preferably: 200～800 parts of kurarinones, 5～350 parts of ganoderans, 50～800 parts of Radix Astragali total saponinss; The best is: 400 parts of kurarinones, 15～250 parts of ganoderans, 100～400 parts of Radix Astragali total saponinss.

In the said ratio, the content of polysaccharide is not less than 50% in the ganoderan, preferably is not less than 80%; The content of polysaccharide is not less than 30% in the astragalus polysaccharides, preferably is not less than 50%; The content of total saponins is not less than 30% in the Radix Astragali total saponins, preferably is not less than 50%, and the content of astragaloside wherein is not less than 1.0%, preferably is not less than 2.0%.Ganoderan, astragalus polysaccharides and Radix Astragali total saponins all can be made by oneself with reference to preceding method.

Pharmaceutical composition of the present invention have protect the liver, effect such as antiviral, raising immunity, antiinflammatory, can be used for the treatment of acute, chronic hepatitis and active liver cirrhosis.

The present invention also further the claimed aforementioned pharmaceutical compositions that comprises as the essential active component and the pharmaceutical composition of pharmaceutically acceptable carrier, for clinically or pharmaceutically acceptable arbitrary dosage form; Can parenteral, mode such as oral, percutaneous dosing is applied to the patient who needs this treatment, is preferably oral formulations, injection.

When being used for parenteral, can be made into injection.Injection means the intravital solution of confession injection, emulsion or the suspension that medicine is made and supplies to face with preceding preparation or be diluted to solution or the sterile preparation of the powder of suspension or concentrated solution that injection can be divided into injection, injectable sterile powder and concentrated solution for injection.Injection means that the confession that medicine is made is injected into sterile solution type injection, emulsion-type injection or the suspension type injection of using in the body, can be used for intramuscular injection, intravenous injection, intravenous drip etc.; Its specification has 1ml, 2ml, 5ml, 10ml, 20ml, 50ml, 100ml, 200ml, 250ml, 500ml etc., and wherein large volume (generally the being not less than 100ml) injection of using for intravenous drip also claims venous transfusion.Injectable sterile powder means that confession that medicine is made is faced with the suitable sterile solution of preceding usefulness and is mixed with settled solution or the evenly sterilized powder or the aseptic block of suspension, available suitable solvent for injection preparation back injection, also available venous transfusion preparation posterior vein instils; Sterilized powder makes with solvent crystallization, spray drying method or freeze-drying etc.Concentrated solution for injection means that confession that medicine is made faces the aseptic concentrated solution of using for intravenous drip with preceding dilution.

When making injection, can adopt the conventional method production in the existing pharmaceutical field, optional use solvent or non-aqueous solvent.The most frequently used aqueous solvent is a water for injection, also available 0.9% sodium chloride solution or other suitable aqueous solutions; Non-aqueous solvent commonly used is a vegetable oil, is mainly the injection soybean oil, and other also have the aqueous solution of ethanol, propylene glycol, Polyethylene Glycol etc.During the preparation injection, can not add additives, also can add suitable additives, as osmotic pressure regulator, pH value regulator, solubilizing agent, filler, antioxidant, antibacterial, emulsifying agent, suspending agent etc. according to the character of medicine.Osmotic pressure regulator commonly used comprises sodium chloride, glucose, potassium chloride, magnesium chloride, calcium chloride, sorbitol etc., preferred sodium chloride or glucose; PH value regulator commonly used comprises acetic acid-sodium acetate, lactic acid, citric acid-sodium citrate, sodium bicarbonate-sodium carbonate etc.; Solubilizing agent commonly used comprises polyoxyethylene sorbitan monoleate, propylene glycol, lecithin, polyoxyethylene castor oil etc.; Filler commonly used comprises lactose, mannitol, sorbitol, dextran etc.; Antioxidant commonly used has sodium sulfite, sodium sulfite, sodium pyrosulfite etc.; Antibacterial commonly used is phenol, cresol, chlorobutanol etc.Injection container commonly used has glass ampule, vial, plastic ampoule, plastic bottle etc.

Be used for when oral, can be made into conventional solid preparation, as tablet, capsule, pill, granule etc.; Also can be made into oral liquid, as oral solution, oral suspensions, syrup etc.Tablet means disk shape or the special-shaped flaky solid preparation that medicine and the auxiliary materials and mixing compacting that suits form, based on oral ordinary tablet, other has buccal tablet, Sublingual tablet, mouth paster, chewable tablet, dispersible tablet, fuse, effervescent tablet, slow releasing tablet, controlled release tablet and enteric coatel tablets etc.Capsule means medicine or is added with the adjuvant filling in Capsules or be sealed in solid preparation in the soft capsule material, according to its dissolving and release characteristics, can be divided into hard capsule (being commonly referred to as capsule), soft capsule (soft gelatin capsule), slow releasing capsule, controlled release capsule and enteric coated capsule etc.Pill means medicine and suitable adjuvant uniform mixing, and the spherical or near-spherical solid preparation so that proper method is made comprises drop pill, sugar pill, piller etc.Granule means that medicine and suitable adjuvant make the dried particles shape preparation with certain particle size, can be divided into soluble particles (being commonly referred to as granule), mix suspension grain, effervescent granule, enteric coated particles, slow-releasing granules and controlled release granule etc.Oral solution means that medicine dissolution makes for oral supernatant liquid preparation in suitable solvent.Oral suspensions means the slightly solubility solid drugs, is dispersed in the liquid medium, makes for oral suspension body preparation, also comprises dry suspension or dense suspension.Syrup means the dense aqueous sucrose solution that contains medicine.

When making oral formulations, can add suitable filler, binding agent, disintegrating agent, lubricant etc.Filler commonly used comprises starch, Icing Sugar, calcium phosphate, calcium sulfate two water things, dextrin, microcrystalline Cellulose, lactose, pregelatinized Starch, mannitol etc.; Typical binders comprises sodium carboxymethyl cellulose, PVP-K30, hydroxypropyl cellulose, starch slurry, methylcellulose, ethyl cellulose, hypromellose, gelling starch etc.; Disintegrating agent commonly used comprises dried starch, polyvinylpolypyrrolidone, cross-linking sodium carboxymethyl cellulose, carboxymethyl starch sodium, low-substituted hydroxypropyl cellulose etc.; Conventional lubricants comprises magnesium stearate, Pulvis Talci, sodium lauryl sulphate, micropowder silica gel etc.

The invention has the advantages that:

1, provides a kind of new preparation to treat the Chinese medicine compound of the medicine of hepatic disease, satisfied urgent clinical needs.

2, pharmaceutical composition of the present invention has been carried out pharmaceutical research, the result shows: pharmaceutical composition of the present invention can extremely significantly reduce the ALT and the AST of immunologic liver injury mice serum, and the mouse immune liver damage is had significant protective effect; DHB all there is utmost point significant inhibitory effect; Chronic hepatitis there is good improvement effect; Hepatic fibrosis rats serum TG F β 1 level to tetrachloro-methane induction significantly reduces, and the liver tissues of rats area of collagen obviously reduces.It is individually dosed that its effect obviously is better than Ganoderma, the Radix Astragali and kurarinone, and the prompting three has the effect of Synergistic.

3, irritate of the research of harmonization of the stomach drug administration by injection by the different proportionings of the present composition, filtered out the optimum ratio of the present composition the protective effect of mouse immune liver damage.

4, the Ganoderma in the present composition and the Radix Astragali both can have been fed intake by Ganoderma, the Radix Astragali and make, and also can directly be fed intake by ganoderan, astragalus polysaccharides or Radix Astragali total saponins, and its preparation method is provided, and met the needs of large-scale production.

5, the main content of effective of present composition extract is limited respectively, be convenient to control the quality of product.

6, the present composition has been carried out acute toxicity testing, the result shows that present composition toxicity is little, and safety range is big.

7, the stability experiment result that pharmaceutical composition of the present invention is carried out shows that every index is all more stable, has guaranteed safety of clinical administration.

Below routine by experiment beneficial effect of further setting forth medicine of the present invention.The compositions of kurarinone, ganoderan and astragalus polysaccharides hereinafter to be referred as The KLH composition I, the compositions of kurarinone, ganoderan and Radix Astragali total saponins hereinafter to be referred as The KLH group Compound IIUsed ganoderan is taken from embodiment 1 in the experimental example, and astragalus polysaccharides is taken from embodiment 2, and Radix Astragali total saponins is taken from embodiment 3.

Experimental example 1 KLH compositions drug combination pharmacodynamic study

Test sample:

0.9% normal saline, self-control;

Chemical pure CCl ₄, Chengdu associating chemical reagent institute;

Matrine Injection, 2ml: 0.2g, Tianjin Biochemical Pharmaceutical Factory;

The Ganoderma injection, self-control, 5ml contains ganoderan 91.8mg (being equivalent to Ganoderma 6g);

Astragalin injection, self-control, 5ml contains astragalus polysaccharides 214.0mg (being equivalent to Radix Astragali 20g);

The Radix Astragali total saponins injection, self-control, 5ml contains Radix Astragali total saponins 212.0mg (being equivalent to Radix Astragali 20g);

The different proportionings of KLH composition I injection (kurarinone+Ganoderma+Radix Astragali), self-control, preparation method is with reference to embodiment 4;

The different proportionings of KLH composition I I injection (kurarinone+Ganoderma+Radix Astragali), self-control, preparation method is with reference to embodiment 4.

Reagent: bacillus calmette-guerin vaccine (BCG), Shanghai Vaccine and Serum Institute; Lipopolysaccharide (LPS), Sigma company.

Animal subject: ICR kind mice, body weight 16～20g, male and female half and half.

Experimental technique: get 240 of mices, mice is divided into normal saline matched group, model group, kurarinone group, Ganoderma group, astragalus polysaccharides group, Radix Astragali total saponins group, the different proportioning groups with KLH composition I I of the different proportioning groups of KLH composition I, 10 every group at random.Except that the every caudal vein injecting normal saline of matched group 0.2ml, other is respectively organized every caudal vein injection 0.2ml bacillus calmette-guerin vaccine and (contains 5 * 10 ⁷Viable bacteria).Play begin treatment next day, matched group and model group mice intraperitoneal injection of saline every day 0.5ml, other respectively organizes the lumbar injection relative medicine, and dosage sees Table 1.After 12 days, except that matched group gave a normal saline 0.2ml/ tail vein injection, all the other each Mus by tail vein injection lipopolysaccharide 0.2ml/7.5 μ g/ only.After 12 hours, eye socket is got blood, and conventional separation of serum is used IFCC recommendation method and measured serum glutamic pyruvic transminase (ALT) and glutamic oxaloacetic transaminase, GOT (AST) activity.

Table 1 KLH compositions is to the influence of immunologic liver injury mice serum ALT and AST (n=10)

Group	Proportioning (mg+g+g)	Dosage (mg/kg)	ALT	AST
Group	Proportioning (mg+g+g)	Dosage (mg/kg)	ALT	AST	Normal saline matched group model group kurarinone group Ganoderma group astragalus polysaccharides group Radix Astragali total saponins group	- - - - - -	- - 90 90 90 90	40.56±8.91 301.34±50.12 ^ΔΔ 198.76±35.34 ^ 214.67±40.16 ^ 235.11±48.86 ^ 237.91±49.32 ^	171.67±29.58 517.38±146.43 ^ΔΔ 386.54±110.35 ^ 402.56±117.34 ^ 470.12±126.54 ^ 468.93±126.79 ^
KLH composition I group (kurarinone+ganoderan+astragalus polysaccharides)	200+1+10 200+6+20 200+12+40 400+1+10 400+6+20 400+12+40 800+1+10 800+6+20 800+12+40	90 90 90 90 90 90 90 90 90	155.34±26.13 ^abc 140.16±23.24 ^abc 136.38±20.12 ^abc 125.47±21.42 ^abc 110.43±18.65 ^abc 118.46±15.37 ^abc 123.43±31.24 ^abc 119.04±29.98 ^abc 117.32±25.64 ^**abc	321.76±110.32 ^abc 318.94±109.78 ^abc 301.56±104.23 ^abc 287.58±94.02 ^abc 226.97±80.16 ^abc 230.14±82.45 ^abc 268.91±92.46 ^abc 259.79±89.94 ^abc 243.14±86.25 ^**abc		- - - - - -	- - 90 90 90 90
		90 90 90 90 90 90 90 90 90			KLH composition I I organizes (kurarinone+ganoderan+Radix Astragali total saponins)	200+1+10 200+6+20 200+12+40 400+1+10 400+6+20 400+12+40 800+1+10 800+6+20 800+12+40	90 90 90 90 90 90 90 90 90	156.38±26.24 ^abd 144.29±24.31 ^abd 134.53±20.26 ^abd 126.51±21.36 ^abd 109.92±18.67 ^abd 117.43±15.45 ^abd 124.42±31.26 ^abd 118.71±29.85 ^abd 115.25±25.51 ^**abd	324.66±110.35 ^abd 319.86±109.69 ^abc 301.62±104.25 ^abc 286.86±94.13 ^abd 227.84±80.21 ^abd 225.24±82.56 ^abd 257.94±92.51 ^abd 262.69±89.85 ^abc 253.16±86.32 ^**abd

Annotate: compare with the normal saline matched group, ^{The Δ Δ}P＜0.01; Compare with model group, ^*P＜0.01; Compare with the kurarinone group, ^aP＜0.01; Compare with the Ganoderma group, ^bP＜0.01; Compare with the astragalus polysaccharides group, ^cP＜0.01; Compare with the Radix Astragali total saponins group, ^dP＜0.01.

Experimental result: as shown in Table 1

1) compare with the normal saline matched group, model group mice serum ALT and AST activity extremely significantly increase (p＜0.01), and the modeling success is described.

2) compare with model group, each administration group all can extremely significantly reduce the ALT and the AST (p＜0.01) of immunologic liver injury mice serum.

3) compare each dosage group injection more remarkable effect (p＜0.01) of KLH composition I with the medicine group with kurarinone group, Ganoderma group, astragalus polysaccharides group list; Compare with the medicine group with kurarinone group, Ganoderma group, Radix Astragali total saponins group list, each dosage group injection more remarkable effect (p＜0.01) of KLH composition I I illustrates that kurarinone, Ganoderma and Radix Astragali three have the effect of Synergistic.

Conclusion: by experimental result as can be known, kurarinone, Ganoderma and Radix Astragali three have share synergistic function.Especially when kurarinone: Ganoderma: effect is best during the Radix Astragali=400: 6000: 20000.

Experimental example 2 KLH compositionss are to immunologic liver injury mice serum ALT and the active influence of AST

Test sample:

0.9% normal saline, self-control;

Chemical pure CCl ₄, Chengdu associating chemical reagent institute;

The kurarinone capsule, 0.1g/ grain, Ningxia Boertaili Pharmaceutical Co., Ltd;

LINGZHI JIAONANG, self-control contains ganoderan 91.8mg/ grain (being equivalent to Ganoderma 6g);

The astragalus polysaccharides capsule, self-control contains astragalus polysaccharides 214.0mg/ grain (being equivalent to Radix Astragali 20g);

The Radix Astragali total saponins capsule, self-control contains Radix Astragali total saponins 212.0mg grain (being equivalent to Radix Astragali 20g);

The different proportionings of KLH composition I capsule (kurarinone+Ganoderma+Radix Astragali), self-control, preparation method is with reference to embodiment 9;

The different proportionings of KLH composition I I capsule (kurarinone+Ganoderma+Radix Astragali), self-control, preparation method is with reference to embodiment 9.

Experimental technique: get 360 of mices, mice is divided into normal saline matched group, model group, kurarinone group, Ganoderma group, astragalus polysaccharides group, Radix Astragali total saponins group, the different proportioning groups with KLH composition I I of the different proportioning groups of KLH composition I, 10 every group at random.Except that the every caudal vein injecting normal saline of normal saline matched group 0.2ml, other is respectively organized every caudal vein injection 0.2ml bacillus calmette-guerin vaccine and (contains 5 * 10 ⁷Viable bacteria).Play begin treatment next day, the every Mus of matched group and model group is irritated stomach normal saline 0.5ml every day, and other respectively organizes gastric infusion relative medicine every day, be diluted to suitable concentration with physiological saline solution before the medicine administration of giving, dosage sees Table 2.After 12 days, except that matched group gave a normal saline 0.2ml/ tail vein injection, all the other each Mus by tail vein injection lipopolysaccharide 0.2ml/7.5 μ g/ only.After 12 hours, eye socket is got blood, and conventional separation of serum is used IFCC recommendation method and measured serum glutamic pyruvic transminase (ALT) and glutamic oxaloacetic transaminase, GOT (AST) activity.

Experimental result: as can be known by table 2 result

3) compare with the medicine group with kurarinone group, Ganoderma group, astragalus polysaccharides group list, each dosage group injection more remarkable effect (p＜0.01) of KLH composition I, compare with the medicine group with kurarinone group, Ganoderma group, Radix Astragali total saponins group list, each dosage group injection more remarkable effect (p＜0.01) of KLH composition I I illustrates that kurarinone, Ganoderma and Radix Astragali three have the effect of Synergistic.

Table 2 KLH compositions is to the influence of immunologic liver injury mice serum ALT and AST (n=10)

Group	Proportioning (mg+g+g)	Dosage (mg/kg)	ALT	AST
Group	Proportioning (mg+g+g)	Dosage (mg/kg)	ALT	AST	Normal saline matched group model group kurarinone group Ganoderma group astragalus polysaccharides group Radix Astragali total saponins group	- - - - - -	- - 90 90 90 90	41.05±8.26 307.25±49.26 ^ΔΔ 202.63±32.51 ^ 226.72±43.28 ^ 240.13±42.66 ^ 235.28±47.35 ^	170.82±26.53 508.42±151.35 ^ΔΔ 391.61±98.58 ^ 411.17±106.51 ^ 475.25±115.32 ^ 472.78±118.27 ^
KLH composition I group (kurarinone+ganoderan+astragalus polysaccharides)	100+0.5+1 100+24+80 200+3+40 200+6+20 200+9+10 400+1+40 400+3+20 400+6+20 400+9+20 400+12+10 800+3+40 800+6+20 800+9+10 1600+0.5+1 1600+24+80	90 90 90 90 90 90 90 90 90 90 90 90 90 90 90	157.83±24.32 ^abc 142.56±21.35 ^abc 141.26±25.12 ^abc 135.53±22.24 ^abc 133.25±25.58 ^abc 128.56±23.72 ^abc 121.73±19.52 ^abc 112.38±21.05 ^abc 119.62±18.53 ^abc 124.37±22.65 ^abc 129.38±33.56 ^abc 135.37±30.55 ^abc 132.05±28.37 ^abc 140.15±20.56 ^abc 156.54±27.78 ^**abc	323.96±97.28 ^abc 316.85±118.16 ^abc 313.35±109.23 ^abc 303.71±112.35 ^abc 288.35±76.56 ^abc 277.58±94.02 ^abc 236.66±82.37 ^abc 221.31±78.56 ^abc 235.41±85.58 ^abc 258.57±85.36 ^abc 261.41±80.35 ^abc 283.68±102.53 ^abc 272.58±95.82 ^abc 310.76±106.36 ^abc 321.93±85.89 ^**abc		- - - - - -	- - 90 90 90 90
		90 90 90 90 90 90 90 90 90 90 90 90 90 90 90			KLH composition I I organizes (kurarinone+ganoderan+Radix Astragali total saponins)	100+0.5+1 100+24+80 200+3+40 200+6+20 200+9+10 400+1+40 400+3+20 400+6+20 400+9+20 400+12+10 800+3+40 800+6+20 800+9+10 1600+0.5+1 1600+24+80	90 90 90 90 90 90 90 90 90 90 90 90 90 90 90	153.62±21.54 ^abd 143.91±22.6 ^abd 132.56±20.18 ^abd 131.25±21.52 ^abd 128.51±26.58 ^abd 126.19±30.57 ^abd 113.57±27.58 ^abd 108.45±17.76 ^abd 116.56±19.96 ^abd 123.17±20.61 ^abd 125.51±28.12 ^abd 129.39±20.55 ^abd 129.26±30.23 ^abd 141.32±19.31 ^abd 150.25±18.62 ^**abd	320.62±111.15 ^abd 318.76±106.58 ^abd 303.56±104.37 ^abd 278.66±92.32 ^abd 271.62±88.29 ^abd 262.58±90.53 ^abd 243.34±87.75 ^abd 232.52±85.16 ^abd 241.27±88.25 ^abd 256.39±91.32 ^abd 259.96±93.55 ^abd 251.28±83.26 ^abd 273.75±85.53 ^abd 315.67±112.35 ^abd 319.27±115.52 ^**abd

Experimental example 3 KLH compositions anti-hepatitis virus experimentatioies

Test sample: 0.9% normal saline, self-control;

The injection acyclovir, Zhongnuo Pharmaceutical (Shijiazhuang) Co., Ltd., Shiyao Group;

The kurarinone capsule, 0.1g/ grain, Ningxia Boertaili Pharmaceutical Co., Ltd;

Ganoderma particles, self-control contains ganoderan 91.8mg/ bag (being equivalent to Ganoderma 6g);

Astragalus polysaccharide particles, self-control contains astragalus polysaccharides 214.0mg/ bag (being equivalent to Radix Astragali 20g);

The Radix Astragali total saponins granule, self-control contains Radix Astragali total saponins 212.0mg/ bag (being equivalent to Radix Astragali 20g);

KLH composition I granule (kurarinone+Ganoderma+Radix Astragali 400mg+6g+20g), self-control, preparation method is with reference to embodiment 10;

KLH composition I I granule (kurarinone+Ganoderma+Radix Astragali 400mg+6g+20g), self-control, preparation method is with reference to embodiment 10.

Laboratory animal: commercially available 1 age in days Beijing duck

DHBV (DHB) positive serum, microbiology teaching and research room of Medical Center of Fudan University.

Animal model: Beijing duck is got blood through sufficient intravenous injection 0.2mlDHBV positive serum behind the 7d, separation of serum, and-20 ℃ of preservations are to be checked.

Experimental technique: filter out 72 of the positive ducks that infect successfully, be divided into 12 groups at random, be respectively blank group, positive controls, kurarinone group, Ganoderma group, astragalus polysaccharides group, Radix Astragali total saponins group, the basic, normal, high dosage group of KLH composition I and the basic, normal, high dosage group of KLH composition I I, 6 every group.The blank group is irritated stomach normal saline 20ml/kg every day, every day 2 times, 14d continuously; Administration group gastric infusion relative medicine, be diluted to suitable concentration with physiological saline solution before the medicine administration of giving, every day 2 times, dosage sees the following form, continuously 14d.Positive drug is pressed 100mg/kg with (acyclovir) ACV and is irritated stomach, 2 times/d, 2 weeks of administration as the treatment matched group.Respectively at 1d (T before the medication ₀), the 7th day (T of medication ₇), the 14th day (T of medication ₁₄) and drug withdrawal after the 7th day (P ₇).Get blood from duck lower limb shin vein, separation of serum ,-20 ℃ of preservations are to be checked.

The DHBV-DNA detection method: it is clear to get above-mentioned Sanguis Anas domestica, and every batch with the time point film, measure Sanguis Anas domestica clear in the variation of DHBV-DNA level, press nick translation test kit illustration method, usefulness ³²P labelling DHBV-DNA probe, and make the clear dot blot hybridization of Sanguis Anas domestica, autoradiography diaphragm speckle is measured OD value (the about 490nm of optical filter wavelength) on enzyme mark detector, calculate serum DHBV-DNA density.

Experimental result: as seen by table 3 result

1) with the normal saline matched group relatively, positive control (acyclovir) group after administration the 7th day and the 14th day has significance (p＜0.01) with difference before the administration, but rebound phenomenon is arranged after the drug withdrawal, does not have significance (p＞0.05) with comparing difference before the administration.

2) with the normal saline matched group relatively, four individually dosed group of kurarinone group, Ganoderma group, astragalus polysaccharides group, Radix Astragali total saponins group after administration the 7th day and the 14th day have significance (p＜0.05) with difference before the administration, and do not have rebound phenomenon after the drug withdrawal.

3) with the normal saline matched group relatively, three various dose groups of KLH composition I, three various dose groups of KLH composition I I after administration the 7th day and the 14th day have significance (p＜0.01) with difference before the administration, and no rise phenomenon after the drug withdrawal.

4) compare for individually dosed group with kurarinone group, Ganoderma group, astragalus polysaccharides group, three various dose groups of KLH composition I after administration the 7th day and the 14th day, with the difference before the administration significance (p＜0.05) is arranged, compare for individually dosed group with kurarinone group, Ganoderma group, Radix Astragali total saponins group, three various dose groups of KLH composition I I after administration the 7th day and the 14th day have significance (p＜0.05) with difference before the administration.

Table 3 KLH compositions is to the inhibitory action of DHBV-DNA in the duck body

Group	Dosage (mg/kg)	The OD490 value
		The OD490 value				T ₀	T ₇	T ₁₄	P ₇
		Physiological saline control group positive controls kushenin group glossy ganoderma group astragalus polyose group astragalus root total saponin group	- - 90 90 90 90	2.27±0.58 2.13±0.43 2.21±0.40 2.26±0.48 2.19±0.53 2.31±0.51	2.08±0.47 0.65±0.53 ^** 1.21±0.37 ^* 1.39±0.41 ^* 1.42±0.45 ^* 1.44±0.43 ^*	T ₀	T ₇	T ₁₄	P ₇	2.03±0.49 0.53±0.46 ^** 1.08±0.32 ^* 1.15±0.38 ^* 1.19±0.39 ^* 1.21±0.42 ^*	2.06±0.47 2.02±0.70 1.32±0.36 ^* 1.38±0.42 ^* 1.43±0.44 ^* 1.45±0.46 ^*
Dosage group KLH composition I low dose group in the KLH composition I high dose group KLH composition I	90 70 50		- - 90 90 90 90	2.27±0.58 2.13±0.43 2.21±0.40 2.26±0.48 2.19±0.53 2.31±0.51		2.25±0.29 2.18±0.32 2.13±0.34	0.81±0.23 ^#abc 0.83±0.28 ^#abc 0.90±0.31 ^**#abc	0.79±0.20 ^#abc 0.77±0.25 ^#abc 0.85±0.28 ^**#abc	0.82±0.24 ^#abc 0.85±0.29 ^#abc 0.91±0.33 ^**#abc
	90 70 50	Dosage group KLH composition I I low dose group among the KLH composition I I high dose group KLH composition I I	90 70 50	2.22±0.29 2.17±0.32 2.16±0.34	0.79±0.25 ^#abd 0.82±0.28 ^#abd 0.87±0.32 ^**#abd	2.25±0.29 2.18±0.32 2.13±0.34	0.81±0.23 ^#abc 0.83±0.28 ^#abc 0.90±0.31 ^**#abc	0.79±0.20 ^#abc 0.77±0.25 ^#abc 0.85±0.28 ^**#abc	0.82±0.24 ^#abc 0.85±0.29 ^#abc 0.91±0.33 ^**#abc	0.76±0.24 ^#abd 0.82±0.30 ^#abd 0.86±0.27 ^**#abd	0.76±0.27 ^#abd 0.81±0.31 ^#abd 0.86±0.34 ^**#abd

Annotate: compare with the blank group, ^*P＜0.05, ^*P＜0.01; Compare with positive controls, ^#P＜0.05; Compare with the kurarinone group, ^aP＜0.05; Compare with the Ganoderma group, ^bP＜0.05; Compare with the astragalus polysaccharides group, ^cP＜0.05; Compare with the Radix Astragali total saponins group, ^dP＜0.05.

Conclusion: each administration group more all has significance to the inhibitory action of DHBV-DNA in duck body difference and the normal saline group before the 7th day and the 14th day and the administration after administration, but after the positive controls drug withdrawal rebound phenomenon is arranged.Pharmaceutical composition curative effect of the present invention all is better than single with kurarinone, Ganoderma, astragalus polysaccharides and Radix Astragali total saponins group, and effect is remarkable, and does not have rebound phenomenon, illustrates that kurarinone, Ganoderma and the Radix Astragali three medicines share, and have synergistic function.

The protective action of 4 pairs of rat chronic hepatic injury of experimental example

Laboratory animal: male Wistar rat, Mus 8 weeks of age, body weight (207 ± 21) g

Test sample:

0.9% normal saline, self-control;

Chemical pure CCl ₄, Chengdu associating chemical reagent institute;

The kurarinone capsule, 0.1g/ grain, Ningxia Boertaili Pharmaceutical Co., Ltd;

Ganoderma tablet, self-control contains ganoderan 91.8mg/ sheet (being equivalent to Ganoderma 6g);

The astragalus polysaccharides sheet, self-control contains astragalus polysaccharides 214.0mg/ sheet (being equivalent to Radix Astragali 20g);

The Radix Astragali total saponins sheet, self-control contains Radix Astragali total saponins 212.0mg/ sheet (being equivalent to Radix Astragali 20g);

KLH composition I injection (kurarinone+Ganoderma+Radix Astragali 400mg+6g+20g), self-control, preparation method is with reference to embodiment 8;

KLH composition I I injection (kurarinone+Ganoderma+Radix Astragali 400mg+6g+20g), self-control, preparation method is with reference to embodiment 8.

Experimental technique: 216 rats are divided into normal control group, model group, kurarinone group, Ganoderma group, astragalus polysaccharides group, Radix Astragali total saponins group, each dosage group of KLH composition I and each dosage group of KLH composition I I at random, equal 18 of every group of rat.Except that the normal control group, all the other each groups are all used 50%CCl ₄Thigh subcutaneous injection behind the-olive oil 1mL/kg, 2 times weekly, in totally 12 weeks, cause the rat chronic hepatic injury, irritate stomach simultaneously respectively and give relative medicine, be diluted to suitable concentration with physiological saline solution, every day 1 time before the medicine administration of giving.The normal control group is irritated stomach equivalent normal saline, every day 1 time.Each is organized and puts to death 6 rats at random in the 4th, 9,12 weeks of experiment.

Leaving and taking of specimen: lumbar injection 2.5% pentobarbital solution 50mg/kg opens the abdominal cavity with anesthesia along hunter's line, gets blood from postcava, separation of serum ,-20 ℃ of preservations; Get liver 10% formaldehyde fixed.

Serological index detects: ALT detects with the full-automatic biochemical analyzer.

Statistical procedures: data with

Expression is handled corresponding data with F check and t check.

Experimental result:

Compare with the normal control group, the ALT level in the 4th, 9,12 weeks of model group rat all significantly raises (p＜0.01), and the modeling success is described; Compare with model group, kurarinone group, Ganoderma group, astragalus polysaccharides group, the ALT level in the 4th, 9,12 weeks of Radix Astragali total saponins group rat all obviously reduce (p＜0.05); The ALT level in each the 4th, 9,12 week of dosage group rat of each dosage group of KLH composition I and KLH composition I I all significantly reduces (p＜0.01).

Table 4 KLH compositions is to the protective effect of chronic hepatic injury mice

Group	Dosage (mg/kg)	ALT
		ALT			4 (weeks)	9 (weeks)	12 (weeks)
		Dosage group KLH composition I I high dose group among the high low dose group KLH composition I of the dosage group KLH composition I I low dose group KLH composition I I in the Normal group model group kushenin group glossy ganoderma group astragalus polyose group astragalus root total saponin group KLH composition I low dose group KLH composition I	- - 100 100 100 100 65 80 100 65 80 100	88.70±20.16 272.83±18.66 ^ΔΔ 220.33±53.54 ^* 237.65±51.35 ^* 240.23±48.67 ^* 241.31±46.15 ^* 205.25±51.35 ^abc 198.22±46.53 ^abc 185.63±52.55 ^abc 201.32±48.58 ^abd 193.56±55.13 ^abc 178.37±50.72 ^abd	4 (weeks)	9 (weeks)	12 (weeks)	81.45±19.37 261.00±52.44 ^ΔΔ 203.33±49.06 ^* 221.35±45.26 ^* 233.26±41.36 ^* 236.29±45.66 ^* 182.21±43.62 ^abc 175.16±45.28 ^abc 162.35±51.23 ^abc 178.32±40.06 ^abd 155.53±42.56 ^abc 148.26±41.76 ^abd	84.50±16.57 296.17±43.13 ^ΔΔ 222.50±51.34 ^* 235.65±52.51 ^* 238.66±53.47 ^* 241.07±53.51 ^* 193.36±50.63 ^abc 185.41±48.37 ^abc 172.65±51.35 ^abc 186.85±50.72 ^abd 177.43±51.29 ^abd 165.67±48.93 ^abd

Annotate: compare with the normal control group, ^{The Δ Δ}P＜0.01; Compare with model group, ^*P＜0.05, ^*P＜0.01; Compare with the kurarinone group, ^aP＜0.05; Compare with the Ganoderma group, ^bP＜0.05; Compare with the astragalus polysaccharides group, ^cP＜0.05; Compare with the Radix Astragali total saponins group, ^dP＜0.05.

Conclusion: serum ALT activities is widely used as clinical diagnosis liver inflammatory damage and judges the sensitive indicator of its clinical efficacy.Show that by above result KLH composition I and KLH composition I I can suppress inflammation activity and proliferation of fibrous tissue in the hepatic tissue, thereby to CCl ₄The rat chronic that causes is done damage and is had preventive and therapeutic effect.

The Hepar Mus fibrosis effect of the experimental example 5 KLH compositions Chinese People's Anti-Japanese Military and Political College

Animal subject: healthy male SD rat, 120, body weight 140～160g is divided into 12 groups at random, 10 every group.

Test sample: 0.9% normal saline, self-control

Chemical pure CCl ₄, Chengdu associating chemical reagent institute;

Matrine Injection, 2ml: 0.2g, Tianjin Biochemical Pharmaceutical Factory;

The Radix Astragali total saponins injection, self-control, 5m1 contains Radix Astragali total saponins 212.0mg (being equivalent to Radix Astragali 20g);

KLH composition I injection (kurarinone+Ganoderma+Radix Astragali 400mg+6g+20g), self-control, preparation method is with reference to embodiment 4;

KLH composition I I injection (kurarinone+Ganoderma+Radix Astragali 400mg+6g+20g), self-control, preparation method is with reference to embodiment 4.

Experimental technique: 120 rats are divided into 12 groups at random, every group 10, be respectively blank group, model according to group, kurarinone group, Ganoderma group, astragalus polysaccharides group, Radix Astragali total saponins group, the basic, normal, high dosage group of KLH composition I and the basic, normal, high dosage group of KLH composition I I.Use CCl ₄Hypodermic injection is induced the rat liver fibrosis model, promptly uses 300ml/LCCl ₄Paraffin oil solution is done subcutaneous injection with 3ml/kg, and 2 times weekly, in totally 8 weeks, the blank group gives equivalent normal saline subcutaneous injection.Each administration group intraperitoneal injection in modeling, dosage sees Table 5, CCl ₄The model group modeling gives the normal saline lumbar injection simultaneously, and the blank group gives isopyknic normal saline.Each treated animal is CCl the last time ₄Injection back 48h puts to death, and it is to be tested to get serum regulating liver-QI tissue specimen.

Detection method: serum is monitored TGF β 1 level according to the reagent description with enzyme linked immunosorbent assay (ELISA), the hepatic tissue specimen with neutral formalin solution fix, paraffin embedding, microscope slide with the poly-D-lysine coating is made 5 μ m tissue slices, carries out HE dyeing and the triple collagen stainings of Massion and makes histopathological examination; (immunohistochemistry, IH) method detects Smad 3 protein expression levels, observed result under the optical microscope to immunohistochemistry.

Table 5 is respectively organized the liver tissues of rats check result

Group	Dosage (mg/kg)	TGFβ1(mg/ml)	Histology's integration	Area of collagen (μ m ²)	Smad3 positive rate (%)
Group	Dosage (mg/kg)	TGFβ1(mg/ml)	Histology's integration	Area of collagen (μ m ²)	Smad3 positive rate (%)	Dosage group KLH composition I I low dose group among the dosage group KLH composition I low dose group KLH composition I I high dose group KLH composition I I in the blank group model group kushenin group glossy ganoderma group astragalus polyose group astragalus root total saponin group KLH composition I high dose group KLH composition I	- - 100 100 100 100 100 80 65 100 80 65	15.69±6.34 96.37±16.56 ^## 36.65±8.72 ^* 40.13±8.57 ^* 41.89±8.97 ^* 42.13±9.24 ^* 22.14±6.51 ^abc 25.23±6.76 ^abc 27.14±7.59 ^abc 21.89±6.56 ^abc 25.04±6.84 ^abc 26.95±7.78 ^abc	0 3.6±0.8 ^## 1.9±0.8 ^* 2.2±0.8 ^* 2.4±0.7 ^* 2.3±0.9 ^* 0.8±0.5 ^abc 1.1±0.7 ^abc 1.3±0.9 ^abc 0.7±0.6 ^abd 1.1±0.9 ^abd 1.2±0.8 ^abd	56.33±21.42 690.86±188.71 ^## 99.42±37.25 ^ 110.32±63.24 ^ 126.96±108.35 ^ 125.87±103.54 ^ 64.28±22.51 ^abc 73.25±21.59 ^abc 86.75±24.46 ^abc 62.91±21.26 ^abd 72.88±23.13 ^abd 86.47±25.58 ^abd	0.86±0.03 9.56±1.35 ^## 4.33±1.65 ^* 5.53±1.15 ^* 6.04±1.24 ^* 6.13±1.68 ^* 1.01±0.86 ^abc 1.73±1.02 ^abc 2.21±1.12 ^abc 1.06±1.07 ^abd 1.65±1.14 ^abd 2.19±1.23 ^abd

Annotate: compare with the blank group, ^##P＜0.01; Compare with model group, ^*P＜0.05, ^*P＜0.01; Compare with the kurarinone group, ^aP＜0.05; Compare with the Ganoderma group, ^bP＜0.05; Compare with the astragalus polysaccharides group, ^cP＜0.05; Compare with the Radix Astragali total saponins group, ^dP＜0.05.

Experimental result: as seen by table 5 result

1) compare with the blank group, model group rat blood serum TGF β 1 level significantly raises (p＜0.01), illustrates that the forming process of hepatic fibrosis is attended by TGF β 1 synthetic remarkable increase, thereby promotes the synthetic and deposition of liver collagen fiber; The extensive steatosis of visible model group rat hepatocytes, necrosis (p＜0.01) under the HE dyeing light microscopic, Masson dyeing sees that blue collagen fiber obviously increase (p＜0.01); Immuning tissue's detection shows that model group liver tissues of rats Smad3 protein expression obviously strengthens (p＜0.01); Above result all illustrates the modeling success.

2) compare with model group, kurarinone group, Ganoderma group, astragalus polysaccharides group, four individually dosed group TGF β 1 level of Radix Astragali total saponins group, histology's integration and hepatic tissue Smad3 protein expression all obviously reduce (p＜0.05), and area of collagen significantly reduces (p＜0.01); Each dosage group of KLH composition I, each dosage group TGF β 1 level of KLH composition I I, histology's integration, hepatic tissue Smad3 protein expression and area of collagen all significantly reduce (p＜0.01).

3) compare for individually dosed group with kurarinone group, Ganoderma group, astragalus polysaccharides group, each dosage group TGF β 1 level of KLH composition I, histology's integration, hepatic tissue Smad3 protein expression and area of collagen all obviously reduce (p＜0.05), compare for individually dosed group with kurarinone group, Ganoderma group, Radix Astragali total saponins group, each dosage group TGF β 1 level of KLH composition I I, histology's integration, hepatic tissue Smad3 protein expression and area of collagen all obviously reduce (p＜0.05).

Conclusion: compare with model group, the preventative-therapeutic rat blood serum TGF of each administration group β 1 level significantly reduces, and the liver tissues of rats area of collagen obviously reduces, and the Smad3 The positive expression rate obviously reduces; And the effect of each dosage group of KLH composition I, each dosage group of KLH composition I I is better than four individually dosed group of kurarinone group, Ganoderma group, astragalus polysaccharides group, Radix Astragali total saponins group.

Experimental example 6 injected in mice administration acute toxicity testings

(1) experimental technique

Test sample: KLH composite injection I, dosage form: liquid drugs injection, specification: 10ml derives from embodiment 4;

KLH composite injection II, dosage form: liquid drugs injection, specification: 10ml derives from embodiment 4.

Animal subject: mice, each 5 of every group of male and female, male body weight 25～28g, female body weight 21～24g.

Route of administration: intravenous injection, lumbar injection.

Observe special project: death toll, general state, body weight, cut open inspection, half lethal dose.

(2) experimental result

Require to carry out prerun according to acute toxicity testing, lumbar injection and intravenous injection two route of administration all can't be measured the median lethal dose(LD 50) of medicine, also do not see tangible toxic reaction, so carry out a day maximum dosage-feeding experiment.Dosage: tail vein injection 0.2ml/10g, lumbar injection 0.2ml/10g, 2 times on the one.

Death toll: do not occur dead.

General state: no abnormality seen changes.

Body weight: in administration preceding 1 day, administration day, measured in 2,4,6,8,10,12,14 days after the administration; No abnormality seen changes.

Cut open inspection: the heart, liver, lung, kidney etc. organize no abnormality seen to change.

(3) conclusion

Occur death in this experiment, KLH composite injection I, KLH composite injection II are 0.4ml/10g to the maximum tolerated dose of male and female mouse vein and intraperitoneal injection, are equivalent to 100 times of maximum consumption 20ml of the 50kg body weight day for human beings.Show this product low toxicity, safe.

Experimental example 7 KLH composite injection stability experiments

Investigation project: character, pH value, clarity, related substance, sign content; And at accelerated tests 6 months and the aseptic and pyrogen test of long-term experiment end of term increase.

1, influence factor's experiment

The strong illumination experiment: get test sample, putting illumination is interior the placement 10 days of lighting box of 4500Lx.

High temperature experiment: get test sample, place respectively under 40 ℃, the 60 ℃ conditions and placed 10 days.

Low temperature test: get test sample, in 4 ℃ of refrigerators, placed 10 days.

Above-mentioned experiment was respectively at the 5th, 10 day sampling and measuring.Relatively test every index after the character, and with result and comparison in 0 day.

The result: placed 10 days under the illumination 4500Lx condition, except that related substance slightly raise, all other indexs had no significant change.Placed 10 days under 60 ℃ of conditions of high temperature, outward appearance becomes faint yellow clear liquid, indicates content and descends, and related substance slightly raises.Placed 10 days under 40 ℃ of high temperature, 4 ℃ of conditions of low temperature, every index does not have significant change.

2, accelerated tests

Method: put under the condition of 40 ℃ ± 2 ℃ of temperature, relative humidity 75% ± 5% and placed 6 months.Respectively at taking a sample 1st month, 2 months, 3 months, 6 the end of month, relatively after the outward appearance, test every index at experimental session, with result and comparison in 0 month; And at 6 aseptic and pyrogen tests of increase at the end of month.

Result: placed 6 months under the condition of 40 ℃ ± 2 ℃ of temperature, relative humidity 75% ± 5%, removing related substance slightly increases, and outside sign content slightly descended, all other indexs had no significant change, at 6 the end of month of accelerated tests, pyrogen, sterility test are all up to specification.

3, long-term experiment

Method: put under the condition of 25 ℃ ± 2 ℃ of temperature, relative humidity 60% ± 10% and placed 18 months.Respectively at 3rd month, 6 months, 9 months, 12 months, 18 months, relatively after the outward appearance, test every index, with result and comparison in 0 month; And at 18 aseptic and pyrogen tests of increase at the end of month.

The result: placed under the condition of 25 ℃ ± 2 ℃ of temperature, relative humidity 60% ± 10% 18 months, every index has no significant change, and at 18 the end of month of long-term experiment, pyrogen, sterility test are all up to specification.

Conclusion: reached a conclusion by above-mentioned investigation result, in every experiment, medicine composition injection of the present invention is more stable.

[specific embodiment]

Come by the following examples further to set forth preparation of drug combination method of the present invention, but this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following embodiment.All technology that realizes based on foregoing of the present invention all belong to scope of the present invention.The adjuvant of each dosage form can be replaced with acceptable accessories in following examples, perhaps reduces, increases.Ganoderan in following examples 4～11 is taken from embodiment 1, and astragalus polysaccharides is taken from embodiment 2, and Radix Astragali total saponins is taken from embodiment 3.

The preparation of embodiment 1 ganoderan

Get the Ganoderma medical material, be ground into coarse grain, add the moistening of 4 times of amounts of water and spend the night, decoct next day three times, add 12 times of water gagings for the first time and decocted 3 hours, add 10 times of water gagings for the second time and decocted 2 hours, add 10 times of water gagings for the third time and decocted 1 hour, collecting decoction filters, it is 1.10～1.15 that filtrate is concentrated into relative density, filters, and filtrate is removed albumen with the Sevage method, filter, it is 60% to containing the alcohol amount that filtrate adds ethanol, stirs evenly, cold preservation was left standstill 24 hours, filtered, and collected filter cake, add an amount of water stirring and dissolving, filter, it is 85% to containing the alcohol amount that filtrate adds ethanol, stir evenly, cold preservation was left standstill 24 hours, filtered, collect filter cake, add proper amount of acetone, the washing with alcohol cyclic washing repeatedly, sucking filtration, precipitation adds suitable quantity of water makes dissolving, ultrafiltration is collected molecular weight greater than 50000 daltonian parts, concentrates, vacuum drying, promptly.

Differentiate: get this product 50mg, add ethanol 25ml, reflux 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution.Other gets Ganoderma control medicinal material 2g, shines medical material solution in pairs with legal system.According to the thin layer chromatography experiment, draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel G plate, upper solution with petroleum ether (60～90 ℃)-Ethyl formate-formic acid (15: 5: 1) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

Assay: it is an amount of that the preparation precision of reference substance solution takes by weighing 105 ℃ of glucose reference substances that are dried to constant weight, adds water and make the solution that every 1ml contains 0.1mg, promptly.

The preparation of standard curve is quiet close absorption reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, 1.2ml respectively, put in the 10ml tool plug test tube, add water to 2.0ml, accurate sulphuric acid anthrone solution (the quiet close anthrone 0.1g that takes by weighing that adds, add 80% sulfuric acid solution 100ml and make dissolving, shake up) 6ml, shake up, put in the water-bath and heated 15 minutes, take out, put into water-bath cooling 15 minutes, with the corresponding solvent is blank, measures absorbance under 625nm, is vertical coordinate with the absorbance, concentration is abscissa, the drawing standard curve.

This product 20mg is got in the preparation of need testing solution, puts in the 50ml measuring bottle, is dissolved in water, and is diluted to scale, shakes up promptly.

The accurate need testing solution 2ml that draws of algoscopy puts in the 10ml tool plug test tube, and assay method under the sighting target directrix curve is measured absorbance in accordance with the law, from the weight that standard curve is read the glucose of need testing solution, calculates, promptly.

According to above-mentioned technology, make ganoderan three batch samples, its yield and content see the following form.

Table 6 ganoderan yield and assay result

Batch	Medical material weight (kg)	Ganoderan weight (kg)	Ganoderan yield (%)	Polyoses content (%)
Batch	Medical material weight (kg)	Ganoderan weight (kg)	Ganoderan yield (%)	Polyoses content (%)	123 is average	60 60 60 60	0.75 0.92 1.08 0.92	1.25 1.53 1.80 1.53	82.4 83.2 86.4 84.0

The preparation of embodiment 2 astragalus polysaccharidess

Differentiate: (1) gets the about 0.2g of this product, after adding water 5ml dissolving, add 5 of alkaline cupric tartrate test solutions, heating promptly produces red precipitate, cooling, filter, get filtrate add 1 of hydrochloric acid make acid, heating in water bath 10 minutes, put cold, regulate pH value to neutral, add alkaline cupric tartrate test solution 0.5ml, heating in water bath promptly produces red copper oxidule precipitation.

(2) get the about 0.2g of this product, add water 2ml dissolving after, add 5% alpha-Naphthol alcoholic solution 0.5ml and shake up, slowly add sulphuric acid 3ml, two liquid level intersection displaing amaranth rings.

Assay: the preparation of standard solution takes by weighing the glucose 100mg that is dried to constant weight through 105 ℃, and accurate the title decides, and puts in the 100ml measuring bottle, is dissolved in water, and is diluted to scale, shakes up.Precision is measured 10ml, puts in the 100ml measuring bottle, adds water to scale, shakes up, and is standby.

Standard curve drafting precision is measured totally 6 parts of standard solution 0.1ml～0.6ml, put respectively in the 25ml measuring bottle, add water to 2.0ml, and add phenol solution and (get phenol 300g, aluminium flake 0.3g, sodium bicarbonate 0.15g, mix distillation, collect 182 ℃ of fractions, be mixed with 5% aqueous solution) 1.0ml, shake up, drip concentrated sulphuric acid 5.0ml rapidly, shake up, place 5min, put and heat 15min in the water-bath, take out, be cooled to room temperature rapidly, in addition with the same operation repetitive of 2.0ml water as blank, measure absorption value at 490nm wavelength place, calculate regression equation.

Assay method is got Radix Astragali extract 0.5g and is put in the 25ml measuring bottle, and the method under the sighting target directrix curve drafting item is measured absorption value, according to the content of regression equation calculation polysaccharide from " adding water to 2.0ml " in accordance with the law.

According to above-mentioned technology, make Radix Astragali extract three batch samples, its yield and content see the following form.

The yield of table 7 astragalus polysaccharides and assay result

Batch	Medical material weight (kg)	Astragalus polysaccharides weight (kg)	Astragalus polysaccharides yield (%)	Polyoses content (%)
Batch	Medical material weight (kg)	Astragalus polysaccharides weight (kg)	Astragalus polysaccharides yield (%)	Polyoses content (%)	123 is average	200 200 200 200	1.70 2.08 2.64 2.14	0.85 1.04 1.32 1.07	68.9 72.3 56.5 65.9

The preparation of embodiment 3 Radix Astragali total saponinss

Get the Radix Astragali, decoct with water three times, each 1.5 hours, collecting decoction filters, and it is 1.20～1.25 (60 ℃) that filtrate is concentrated into relative density, handle 2 times with ethanol precipitation, containing the alcohol amount in the solution for the first time is 75%, is 85% for the second time, each all cold preservation is placed, and reclaims ethanol and be concentrated into every 1ml to be equivalent to crude drug 10g, is diluted to every 1ml with water for injection and is equivalent to crude drug 1g, cold preservation was placed 12 hours, filter, filtrate decompression concentrates, vacuum drying, promptly.

Make three batches of Radix Astragali total saponinss respectively, extract yield and content results see the following form 6.

Differentiate one: get this product 0.01g, add methanol 20ml, reflux 1 hour, filter, filtrate is added on neutral alumina post (100～120 orders, 5g, on the internal diameter 10～15mm), with 40% methanol 100ml eluting, collect eluent, evaporate to dryness, residue adds water 30ml makes dissolving, extract 2 times with water saturated n-butyl alcohol jolting, each 20ml merges n-butyl alcohol liquid; Wash each 20ml with water 2 times; Discard water liquid, n-butyl alcohol liquid evaporate to dryness, residue add methanol 0.5ml makes dissolving, as need testing solution.Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution.Drawing each 2 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, is developing solvent with lower floor's solution of chloroform-methanol-water (13: 7: 2), launch, take out airing, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, daylight shows down identical sepia speckle; Ultra-violet lamp (365nm) shows identical orange-yellow fluorescence speckle down.

Differentiate two: get this product 0.01g, add ethanol 30ml, reflux 20 minutes, filter, filtrate adds 0.3% sodium hydroxide solution 15ml makes dissolving, filters, and filtrate is regulated pH value to 5～6 with dilute hydrochloric acid, extract with ethyl acetate 15ml jolting, divide and get acetic acid ethyl fluid, filter the filtrate evaporate to dryness with the filter paper that is covered with anhydrous sodium sulfate, residue adds ethyl acetate 1ml makes dissolving, as need testing solution.Other gets Radix Astragali control medicinal material 2g, shines medical material solution in pairs with legal system.Draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate,, launch as developing solvent with chloroform-methanol (10: 1), take out, airing is put in the ammonia steam and is inspected under the smoked rearmounted ultra-violet lamp (365nm).In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color.

Assay: the assay of total saponins

The preparation precision of reference substance solution takes by weighing the about 10mg of astragaloside reference substance that is dried to constant weight in 105 ℃, puts in the 100ml measuring bottle, adds dissolve with methanol and is diluted to scale, shakes up, and promptly gets (every 1ml contains astragaloside dry product 0.1mg).

This product 0.1g is got in the preparation of need testing solution, and accurate the title decides, and adds water 25ml and makes dissolving, move in the separatory funnel, extract 4 times, each 20ml with water saturated n-butyl alcohol jolting, merge n-butyl alcohol liquid, with the saturated water washing twice of n-butyl alcohol, each 10ml, discard water liquid, n-butyl alcohol evaporate to dryness to the water-bath, residue adds dissolve with methanol, move in the 25ml measuring bottle, and add methanol and be diluted to scale, shake up, filter, get subsequent filtrate, promptly.

The algoscopy precision is measured reference substance solution, each 1ml of need testing solution, puts the 25ml nessler colorimetric tube, puts evaporate to dryness in the water-bath, put coldly, add freshly prepared 5% vanillin-glacial acetic acid solution 0.4ml, perchloric acid 1.6ml, shake up, placed 5 minutes, put in the boiling water bath and developed the color 15 minutes, take out, put immediately and be cooled to room temperature in the ice bath, add the 8ml glacial acetic acid, shake up, measure absorbance at 538nm wavelength place, calculate, promptly.

The assay of astragaloside

Chromatographic condition and system suitability experiment are filler with the octadecyl silane; With acetonitrile-water (32: 68) is mobile phase; Evaporative light scattering detector.Number of theoretical plate calculates with the astragaloside peak and is not less than 4000.

It is an amount of that the preparation precision of reference substance solution takes by weighing the astragaloside reference substance, adds methanol and make the solution that every 1ml contains 0.5mg, promptly.

The preparation precision of need testing solution takes by weighing this product 0.04g, and accurate the title decides, and puts in the apparatus,Soxhlet's, add methanol 40ml, merceration spends the night, and it is an amount of to add methanol again, reflux 4 hours, extracting solution reclaim solvent and are concentrated into driedly, and residue adds water 10ml, slight fever makes dissolving, extracts 4 times with water saturated n-butyl alcohol jolting, each 40ml, merge n-butyl alcohol liquid, use ammonia solution thorough washing 2 times, each 40ml, discard ammoniacal liquor, n-butyl alcohol liquid evaporate to dryness, residue add water 5ml makes dissolving, put cold, by D101 type macroporous adsorptive resins (internal diameter 1.5cm, long 12cm), with water 50ml eluting, discard water liquid, reuse 40% ethanol 30ml eluting, discard eluent, continue with 70% ethanol 80ml eluting, collect eluent, evaporate to dryness, with dissolve with methanol and be transferred in the 5ml measuring bottle, add methanol and be diluted to scale, shake up, promptly.

Accurate respectively reference substance solution 10 μ l, 20 μ l and the need testing solution 20 μ l of drawing of algoscopy inject chromatograph of liquid, measure, and calculate with 2 logarithmic equations of external standard, promptly.

The yield of table 8 Radix Astragali total saponins and assay result

Batch	Medical material (kg)	Total saponins weight (kg)	Total saponins yield (%)	Total saponin content (%)	Astragaloside content (%)
Batch	Medical material (kg)	Total saponins weight (kg)	Total saponins yield (%)	Total saponin content (%)	Astragaloside content (%)	123 is average	200 200 200 200	1.68 2.04 2.64 2.12	0.84 1.02 1.32 1.06	65.4 60.2 58.3 61.3	3.2 3.0 2.5 2.9

The preparation of embodiment 4 KLH compositions aqueous injection

KLH composition I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Astragalus polysaccharides 214.0g (being equivalent to Radix Astragali 20kg)

Polyoxyethylene sorbitan monoleate 20g

Water for injection adds to 10000ml

Prepare 1000 altogether

KLH composition I prescription 2:

Kurarinone 400.0g

Ganoderan 45.9g (being equivalent to Ganoderma 3kg)

Astragalus polysaccharides 428.0g (being equivalent to Radix Astragali 40kg)

Polyoxyethylene sorbitan monoleate 20g

Water for injection adds to 10000ml

Prepare 1000 altogether

KLH composition I I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Radix Astragali total saponins 212.0g (being equivalent to Radix Astragali 20kg)

Polyoxyethylene sorbitan monoleate 50g

Water for injection adds to 10000ml

Prepare 1000 altogether

KLH composition I I prescription 2:

Kurarinone 400.0g

Ganoderan 137.7g (being equivalent to Ganoderma 9kg)

Radix Astragali total saponins 106.0g (being equivalent to Radix Astragali 10kg)

Polyoxyethylene sorbitan monoleate 50g

Water for injection adds to 10000ml

Prepare 1000 altogether

Preparation technology:

Carry and handle the previous day such as pipeline that dosing uses and container etc., face with the fresh water for injection flushing of preceding reuse; Ganoderan is added in the water for injection of dosing amount 30% the heated and stirred dissolving fully; Kurarinone and astragalus polysaccharides (or Radix Astragali total saponins) are added a small amount of water for injection, add the polyoxyethylene sorbitan monoleate heated and stirred dissolving of recipe quantity; Merge above-mentioned solution, benefit adds to the full amount of water for injection; The needle-use activated carbon that adds dosing amount 0.1%, heated and stirred 15 minutes; Through sand filtration rod filtering decarbonization.Measure the also pH value of regulator solution; Microporous filter membrane fine straining through 0.45um; Check the clarity of solution, the semi-finished product chemical examination; With the solution sealing by fusing in glass ampule; 100 ℃ of flowing steam sterilizations 30 minutes; While hot sample being put into 0.01% methylene blue solution hunts leak; Lamp inspection, finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 5 KLH composition powder injections

KLH composition I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Astragalus polysaccharides 214.0g (being equivalent to Radix Astragali 20kg)

Polyoxyethylene sorbitan monoleate 20g

Mannitol 400g

Sterile water for injection adds to 5000ml

Prepare 1000 altogether

KLH compound I prescription 2:

Kurarinone 200.0g

Ganoderan 45.9g (being equivalent to Ganoderma 3kg)

Astragalus polysaccharides 428.0g (being equivalent to Radix Astragali 40kg)

Polyoxyethylene sorbitan monoleate 20g

Mannitol 400g

Sterile water for injection adds to 5000ml

Prepare 1000 altogether

KLH composition I I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Polyoxyethylene sorbitan monoleate 20g

Mannitol 400g

Sterile water for injection adds to 5000ml

Prepare 1000 altogether

KLH composition I I prescription 2:

Kurarinone 200.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Polyoxyethylene sorbitan monoleate 20g

Mannitol 400g

Sterile water for injection adds to 5000ml

Prepare 1000 altogether

Preparation technology:

At first vessel that dosing is used and antibiotic glass bottle, plugs etc. carry out aseptic process; Take by weighing former, adjuvant according to recipe quantity; Ganoderan is added in the water for injection of dosing amount 30% the heated and stirred dissolving fully, kurarinone and astragalus polysaccharides (or Radix Astragali total saponins) are added a small amount of water for injection, add the polyoxyethylene sorbitan monoleate heated and stirred dissolving of recipe quantity, merge above-mentioned solution, add sterile water for injection to full dose; The needle-use activated carbon that adds dosing amount 0.1%, heated and stirred 15 minutes; Through sand filtration rod filtering decarbonization.Measure the also pH value of regulator solution; Microporous filter membrane fine straining through 0.22um; Check the clarity of solution, the semi-finished product chemical examination; Be sub-packed in the antibiotic glass bottle half tamponade.Sample is put into the freeze dryer lyophilization.Pre-freeze-45 ℃ 5 hours, low-temperature vacuum drying-45 ℃～0 ℃ 20 hours was warming up to 25 ℃ of vacuum dryings 3 hours then; Lyophilizing finishes, and lid is rolled in tamponade; Finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 6 KLH compositions sodium chloride transfusion

KLH composition I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Astragalus polysaccharides 214.0g (being equivalent to Radix Astragali 20kg)

Polyoxyethylene sorbitan monoleate 20g

Sodium chloride 900g

Water for injection adds to 100000ml

Prepare 1000 bottles altogether

KLH composition I prescription 2:

Kurarinone 200.0g

Ganoderan 137.7g (being equivalent to Ganoderma 9kg)

Astragalus polysaccharides 107.0g (being equivalent to Radix Astragali 10kg)

Polyoxyethylene sorbitan monoleate 20g

Sodium chloride 900g

Water for injection adds to 100000ml

Prepare 1000 bottles altogether

KLH compositions II prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Polyoxyethylene sorbitan monoleate 20g

Sodium chloride 900g

Water for injection adds to 100000ml

Prepare 1000 bottles altogether

KLH composition I I prescription 2:

Kurarinone 800.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Polyoxyethylene sorbitan monoleate 20g

Sodium chloride 900g

Water for injection adds to 100000ml

Prepare 1000 bottles altogether

Preparation technology:

Handle the previous day such as pipeline that dosing uses and container etc., face with the fresh water for injection flushing of preceding reuse; Ganoderan is added in the water for injection of dosing amount 30% the heated and stirred dissolving fully, kurarinone and astragalus polysaccharides (or Radix Astragali total saponins) are added a small amount of water for injection, the polyoxyethylene sorbitan monoleate heated and stirred dissolving that adds recipe quantity, with sodium chloride fully with an amount of water for injection dissolving, merge above-mentioned solution, benefit adds to the full amount of water for injection; The needle-use activated carbon that adds dosing amount 0.1%, heated and stirred 15 minutes; Through sand filtration rod filtering decarbonization.Measure the also pH value of regulator solution; Microporous filter membrane fine straining through 0.45um; Check the clarity of solution, the semi-finished product chemical examination; Fill is in the infusion bottle of 100ml; 115 ℃ of pressure sterilizings 30 minutes; Lamp inspection, finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 7 KLH compositions glucose infusion liquids

KLH composition I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Astragalus polysaccharides 214.0g (being equivalent to Radix Astragali 20kg)

Polyoxyethylene sorbitan monoleate 20g

Glucose 5000g

Water for injection adds to 100000ml

Prepare 1000 bottles altogether

KLH composition I prescription 2:

Kurarinone 800.0g

Ganoderan 45.9g (being equivalent to Ganoderma 3kg)

Astragalus polysaccharides 428.0g (being equivalent to Radix Astragali 40kg)

Polyoxyethylene sorbitan monoleate 20g

Glucose 5000g

Water for injection adds to 100000ml

Prepare 1000 bottles altogether

KLH composition I I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Polyoxyethylene sorbitan monoleate 20g

Glucose 5000g

Water for injection adds to 100000ml

Prepare 1000 bottles altogether

KLH composition I I prescription 2:

Kurarinone 800.0g

Ganoderan 137.7g (being equivalent to Ganoderma 9kg)

Polyoxyethylene sorbitan monoleate 20g

Glucose 5000g

Water for injection adds to 100000ml

Prepare 1000 bottles altogether

Preparation technology:

Carry and handle the previous day such as pipeline that dosing uses and container etc., face with the fresh water for injection flushing of preceding reuse; Ganoderan is added the heated and stirred dissolving adds a small amount of water for injection with kurarinone and astragalus polysaccharides (or Radix Astragali total saponins) fully in the water for injection of dosing amount 30%, add the polyoxyethylene sorbitan monoleate heated and stirred dissolving of recipe quantity, merge above-mentioned solution; Glucose is complete with the water for injection dissolving of dosing amount 40%, heated and boiled 15 minutes; Merge above-mentioned solution, benefit adds to the full amount of water for injection; The needle-use activated carbon that adds dosing amount 0.1%, heated and stirred 15 minutes; Through sand filtration rod filtering decarbonization.Measure the also pH value of regulator solution; Microporous filter membrane fine straining through 0.45um; Check the clarity of solution, the semi-finished product chemical examination; Fill is in the infusion bottle of 100ml; 115 ℃ of pressure sterilizings 30 minutes; Lamp inspection, finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 8 KLH composition tablets

KLH composition I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Astragalus polysaccharides 214.0g (being equivalent to Radix Astragali 20kg)

Starch 40g

Microcrystalline Cellulose 40g

The 2%HPMC aqueous solution is an amount of

Magnesium stearate 5.0g

Carboxymethylstach sodium 10.0g

. prepare 2000 altogether

KLH composition I prescription 2:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Astragalus polysaccharides 107.0g (being equivalent to Radix Astragali 10kg)

Starch 40g

Microcrystalline Cellulose 40g

The 2%HPMC aqueous solution is an amount of

Magnesium stearate 5.0g

Carboxymethylstach sodium 10.0g

. prepare 2000 altogether

KLH composition I I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Starch 40g

Microcrystalline Cellulose 40g

The 2%HPMC aqueous solution is an amount of

Magnesium stearate 5.0g

Carboxymethylstach sodium 10.0g

Prepare 2000 altogether

KLH composition I I prescription 2:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Radix Astragali total saponins 424.0g (being equivalent to Radix Astragali 40kg)

Starch 40g

Microcrystalline Cellulose 40g

The 2%HPMC aqueous solution is an amount of

Magnesium stearate 5.0g

Carboxymethylstach sodium 10.0g

Prepare 2000 altogether

Preparation technology:

With ganoderan, astragalus polysaccharides (or Radix Astragali total saponins) and standby if the plain pulverize separately of ginseng is crossed 100 mesh sieves; Take by weighing former, adjuvant according to recipe quantity; Hypromellose 2% the aqueous solution made soluble in water is standby; With astragalus polysaccharides (or Radix Astragali total saponins), kurarinone, ganoderan, starch, microcrystalline Cellulose mix homogeneously, adding 2%HPMC aqueous solution is an amount of, stirs, and makes suitable soft material; Cross 20 mesh sieve system granules; Dry under 60 ℃ the condition; Dry good granule adds magnesium stearate and carboxymethylstach sodium, crosses 18 mesh sieve granulate, mix homogeneously; Sampling, the semi-finished product chemical examination; According to the definite sheet weight sheet of chemical examination; Finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 9 KLH composition capsules

KLH composition I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Astragalus polysaccharides 214.0g (being equivalent to Radix Astragali 20kg)

Starch 20.0g

Microcrystalline Cellulose 60.0g

The 2%HPMC aqueous solution is an amount of

Magnesium stearate 5.0g

Prepare 2000 altogether

KLH composition I prescription 2:

Kurarinone 400.0g

Ganoderan 45.9g (being equivalent to Ganoderma 3kg)

Astragalus polysaccharides 107.0g (being equivalent to Radix Astragali 10kg)

Starch 20.0g

Microcrystalline Cellulose 60.0g

The 2%HPMC aqueous solution is an amount of

Magnesium stearate 5.0g

Prepare 2000 altogether

KLH composition I I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Starch 20.0g

Microcrystalline Cellulose 60.0g

The 2%HPMC aqueous solution is an amount of

Magnesium stearate 5.0g

Prepare 2000 altogether

KLH composition I I prescription 2:

Kurarinone 400.0g

Ganoderan 45.9g (being equivalent to Ganoderma 3kg)

Starch 20.0g

Microcrystalline Cellulose 60.0g

The 2%HPMC aqueous solution is an amount of

Magnesium stearate 5.0g

Prepare 2000 altogether

Preparation technology:

It is standby that ganoderan, astragalus polysaccharides (or Radix Astragali total saponins) and kurarinone pulverize separately are crossed 100 mesh sieves; Take by weighing former, adjuvant according to recipe quantity; Hypromellose 2% the aqueous solution made soluble in water is standby; With astragalus polysaccharides (or Radix Astragali total saponins), kurarinone, ganoderan, starch, microcrystalline Cellulose mix homogeneously, adding 2%HPMC aqueous solution is an amount of, stirs, and makes suitable soft material; Cross 20 mesh sieve system granules; Granule is dried under 60 ℃ condition; Dry good granule adds magnesium stearate, crosses 18 mesh sieve granulate, mix homogeneously; Sampling, the semi-finished product chemical examination; The loading amount of determining according to chemical examination incapsulates; Finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 10 KLH composition granules

KLH composition I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Astragalus polysaccharides 214.0g (being equivalent to Radix Astragali 20kg)

Icing Sugar 900.0g

The 2%HPMC50% alcoholic solution is an amount of

Prepare 1000 bags altogether

KLH composition I prescription 2:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Astragalus polysaccharides 107.0g (being equivalent to Radix Astragali 10kg)

Icing Sugar 900.0g

The 2%HPMC50% alcoholic solution is an amount of

Prepare 1000 bags altogether

KLH composition I I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Icing Sugar 900.0g

The 2%HPMC50% alcoholic solution is an amount of

Prepare 1000 bags altogether

KLH composition I I prescription 2:

Kurarinone 400.0g

Ganoderan 137.7g (being equivalent to Ganoderma 9kg)

Icing Sugar 900.0g

The 2%HPMC50% alcoholic solution is an amount of

Prepare 1000 bags altogether

Preparation technology:

It is standby that sucrose was pulverized 100 mesh sieves, and it is standby that ganoderan, astragalus polysaccharides (or Radix Astragali total saponins) and kurarinone pulverize separately are crossed 100 mesh sieves; Take by weighing former, adjuvant according to recipe quantity; With the method mix homogeneously that ganoderan, astragalus polysaccharides (or Radix Astragali total saponins), kurarinone and Icing Sugar progressively increase with equivalent, adding 2%HPMC50% alcoholic solution is an amount of, stirs, and makes suitable soft material, crosses 20 mesh sieve system granules; Granule is dried under 60 ℃ condition; Dried granule is crossed 18 mesh sieve granulate; Sampling, the content of principal agent is determined loading amount in the semi-finished product chemical examination granule; Packing, finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 11 KLH composition oral liquid agent

KLH composition I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Astragalus polysaccharides 214.0g (being equivalent to Radix Astragali 20kg)

Polyoxyethylene sorbitan monoleate 20g

Sodium benzoate 15g

Stevioside 20g

Purified water adds to 10000ml

Prepare 1000 altogether

KLH composition I prescription 2

Kurarinone 400.0g

Ganoderan 137.7g (being equivalent to Ganoderma 9kg)

Astragalus polysaccharides 107.0g (being equivalent to Radix Astragali 10kg)

Polyoxyethylene sorbitan monoleate 20g

Sodium benzoate 15g

Stevioside 20g

Purified water adds to 10000ml

Prepare 1000 altogether

KLH composition I I prescription 1:

Kurarinone 400.0g

Ganoderan 91.8g (being equivalent to Ganoderma 6kg)

Polyoxyethylene sorbitan monoleate 20g

Sodium benzoate 15g

Stevioside 20g

Purified water adds to 10000ml

Prepare 1000 altogether

KLH composition I I prescription 2:

Kurarinone 400.0g

Ganoderan 137.7g (being equivalent to Ganoderma 9kg)

Polyoxyethylene sorbitan monoleate 20g

Sodium benzoate 15g

Stevioside 20g

Purified water adds to 10000ml

Prepare 1000 altogether

Preparation technology:

Ganoderan is added the heated and stirred dissolving adds low amounts of water with kurarinone and astragalus polysaccharides (or Radix Astragali total saponins) fully in the water of dosing amount 30%, add the polyoxyethylene sorbitan monoleate heated and stirred dissolving of recipe quantity, merge above-mentioned solution; Sodium benzoate and stevioside is complete with the water dissolution of dosing amount 20%; Merge above-mentioned two solution, mend and add water to full dose; Cross the filtering with microporous membrane of 0.8um; The semi-finished product chemical examination; Fill, finished product is examined entirely, the packing warehouse-in.

Claims

1. a pharmaceutical composition is characterized in that, makes consisting of of the contained raw materials of effective components medicine of said composition: 100～1600 parts of kurarinones, 500～24000 parts of Ganodermas, 1000～80000 parts of the Radixs Astragali.

2. pharmaceutical composition as claimed in claim 1 is characterized in that, the parts by weight of crude drug are: 200～800 parts of kurarinones, 1000～12000 parts of Ganodermas, 10000～40000 parts of the Radixs Astragali.

3. pharmaceutical composition as claimed in claim 2 is characterized in that, the parts by weight of crude drug are: 400 parts of kurarinones, 3000～9000 parts of Ganodermas, 20000 parts of the Radixs Astragali.

4. as the described arbitrary preparation of drug combination method of claim 1～3, it is characterized in that, the described Ganoderma and the Radix Astragali can singly be carried or mix to obtain through refining and obtain total extract fully with The suitable solvent and method, total extract is made arbitrary preparation with kurarinone and mixing acceptable accessories again, contained main effective ingredient is polysaccharide or polysaccharide and total saponins in the total extract, and the total content of main effective ingredient is not less than 30%.

5. pharmaceutical composition as claimed in claim 1, it is characterized in that, said composition can also be made by following bulk drugs: kurarinone, ganoderan and astragalus polysaccharides or Radix Astragali total saponins, and its weight proportion is: 100～1600 parts of kurarinones, 2.5～700 parts of ganoderans, 5～1600 parts of astragalus polysaccharidess; Perhaps be: 100～1600 parts of kurarinones, 2.5～700 parts of ganoderans, 5～1600 parts of Radix Astragali total saponinss.

6. pharmaceutical composition as claimed in claim 5 is characterized in that, the weight proportion of its crude drug is: 200～800 parts of kurarinones, 5～350 parts of ganoderans, 50～800 parts of astragalus polysaccharidess; Perhaps be: 200～800 parts of kurarinones, 5～350 parts of ganoderans, 50～800 parts of Radix Astragali total saponinss.

7. pharmaceutical composition as claimed in claim 6 is characterized in that, the weight proportion of its crude drug is: 400 parts of kurarinones, 15～250 parts of ganoderans, 100～400 parts of astragalus polysaccharidess; Perhaps be: 400 parts of kurarinones, 15～250 parts of ganoderans, 100～400 parts of Radix Astragali total saponinss.

8. as the described arbitrary pharmaceutical composition of claim 5～7, it is characterized in that the content of polysaccharide is not less than 50% in the described ganoderan; The content of polysaccharide is not less than 30% in the described astragalus polysaccharides; The content of total saponins is not less than 30% in the described Radix Astragali total saponins, and wherein the content of astragaloside is not less than 1%.

9. as claim 1～3,5～7 described arbitrary pharmaceutical compositions, it is characterized in that this pharmaceutical composition can be made clinically any or pharmaceutically acceptable dosage form with mixing acceptable accessories.

10. as claim 1～3, the application of 5～7 described arbitrary pharmaceutical compositions in the medicine of preparation treatment hepatic disease.