CN1969896A - Anti-hepatitis pharmaceutical composition and preparation process thereof - Google Patents

Abstract

The invention discloses a pharmaceutical composition against hepatitis, its preparing process and use, wherein the raw materials of the medicinal constituents include (by weight portions) radix isatidis 200-20000 parts, Hypericum japonicum Thunb. 500-8000 parts, baikal skullcap root glycosides 60-1000 parts, or radix isatidis extract 1-400 parts, Hypericum japonicum Thunb. extract 5-160 parts, baikal skullcap root glycosides 60-1000 parts, the composition can be made into various pharmaceutically acceptable dose forms.

Description

A kind of Antihepatitis medicinal composition and preparation method thereof

1, technical field

The present invention relates to pharmaceutical composition of making by Radix Isatidis or its extract, Herba Hyperici Japonici or its extract and baicalin and its production and use, belong to medical technical field.

2, background technology

Hepatitis is one of common but severe infectious disease, wherein based on viral hepatitis.According to incompletely statistics, have 8～10% to be hepatitis B virus carriers at present in the population of China, and still have every year a considerable amount of new patients to increase.Viral hepatitis can be divided on type: acute hepatitis, chronic hepatitis, hepatitis gravis and gallbladder hepatitis.Though the medicine of treatment hepatopathy has many kinds at present, because the pathogenesis of hepatopathy is not illustrated fully, mostly is and improves liver function, promotes liver cell regeneration, strengthens the medicine of the functions such as detoxification ability of liver.Therefore press for a kind for the treatment of both the principal and secondary aspects of a disease, the medicine of reinforcement and elimination in combination.

Radix Isatidis (Radix Isatidis) is called indigo-blue, Isatis indigotica Fort (Indigofera tinctoria L, Baphicanthus cusia (nees) Brem. Polygonum tinctorium Ait) root, indigo root, is the dry root of Cruciferae Isatis indigotica Fort. platymiscium Isatis indigotica Fort. Isatisindigotica Fort., bitter in the mouth, and cold in nature, GUIXIN, stomach warp have the effect of heat-clearing and toxic substances removing, removing heat from blood sore-throat relieving.Be used for antibiotic, antiviral, antiendotoxin, antiinflammatory, raise immunity etc., be mainly used in treatment influenza, parotitis, epidemic febrile disease heating, send out speckle, anemopyretic cold, swelling throat are mashed, epidemic encephalitis type B, hepatitis etc., be one of traditional anti virus herb, beginning is stated from Shennong's Herbal, and " 1985～2005 years versions of Chinese pharmacopoeia are recorded always.The Radix Isatidis chemical constituent is quite complicated, contains multiple composition,, indirubin indigo as Benzazole compounds etc., the biological alkaline color ammonia of quinoline azole ketone etc., sinigrin compounds, sulfur-bearing compounds, organic acid, base ucleosides guanine etc., amino acids etc.Modern pharmacological research proves that its antibiotic main active is chemical compounds such as color ammonia ketone, the antiviral effective ingredient has purine, pyrimidine, indole etc., the antiendotoxin active substance is an organic acid wherein, and what have immunoregulation effect is the Radix Isatidis polysaccharide, and its index components is the Radix Isatidis total nitrogen.

Herba Hyperici Japonici has another name called Herba Hyperici Japonici, is the herb of Guttiferae plant Herba Hyperici Japonici Hypericum japonicum Thunb..Sweet in the mouth, little hardship, cool in nature, belong to traditional clearing away heat-damp and promoting diuresis, the subduing swelling and detoxicating medicine is mainly used in the treatment infectious hepatitis, dysentery, infantile convulsion, infantile malnutrition, throat moth, acute appendicitis, furuncle and phyma, venom etc.Modern pharmacological research show Herba Hyperici Japonici have antibacterial, protect the liver, suppress tumor and effects such as control cardiovascular system diseases etc., Herba Hyperici Japonici has been extracted and has made injection and be used for the acute and chronic hepatitis of clinical treatment at present, its index components is a Herba Hyperici Japonici total flavones.

Radix Scutellariae is the dry root of labiate Radix Scutellariae Scutellaria baicalensis Georgi, and bitter in the mouth is cold in nature, returns lung, gallbladder, spleen, large intestine, small intestine meridian, have the effect of heat clearing and damp drying, eliminating fire and detoxication, be used for hygropyrexia, fever disease in summer, vomiting and nausea uncomfortable in chest, cough due to lung-heat, diseases such as hyperpyrexia excessive thirst.Baicalin is the effective active composition that extracts in the Radix Scutellariae, be β-maltonic acid-5,6-dihydroxy-4-oxygen-2-phenyl-4H-.alpha.-5:6-benzopyran-7, recorded into the 10th 239 pages of national drug standards chemical drugs provincial standard rising national standards of National Drug Administration (Chinese Pharmacopoeia Commission's volume), wherein regulation contains baicalin (C ₂₁H ₁₈O ₁₁) must not be less than 90.0% (injection); Must not be less than 83.0% (for oral use).Baicalin is an antimicrobial drug, has pharmacological actions such as antibacterial anti-inflammatory, heat-clearing and toxic substances removing, chelated metal ions, calmness, blood pressure lowering, neuroprotective, is mainly used in diseases such as infection, pneumonia, hepatitis, hypertension.Domestic existing 8 families of baicalin raw material listing at present.The structural formula of baicalin is as follows:

Utilize the interaction of Radix Isatidis or its extract, Herba Hyperici Japonici or its extract and baicalin at present, composition of prescription is used for the treatment of the medicine of hepatitis aspect, does not appear in the newspapers as yet.

3, summary of the invention

In order to meet clinical needs, better treat hepatitis, improve the people ' s health level, the invention provides a kind of Antihepatitis medicinal composition and its production and use, mainly be prepared from by Radix Isatidis or its extract, Herba Hyperici Japonici or its extract and baicalin, being used for the treatment of aspects such as acute and chronic hepatitis disease, produced beyond thought effect.

Radix Isatidis in the pharmaceutical composition of the present invention has the effect of heat-clearing and toxic substances removing, can antiinflammatory, and enhancing immunity has therapeutical effect to hepatitis; Herba Hyperici Japonici belongs to traditional clearing away heat-damp and promoting diuresis, the subduing swelling and detoxicating medicine, have antibacterial, effect such as protect the liver; Baicalin is the anti-bacteria and anti-virus medicine, and hepatitis virus is had good inhibitory effect, and three medicine compatibility Synergistics have produced beyond thought effect.

Pharmaceutical composition of the present invention is mainly made by following bulk drugs: 200～20000 parts of Radix Isatidis, 500～8000 parts of Herba Hyperici Japonici, 60～1000 parts of baicalins; Be preferably: 500～15000 parts of Radix Isatidis, 1000～4000 parts of Herba Hyperici Japonici, 125～500 parts of baicalins; Optimum is: 1000～7000 parts of Radix Isatidis, 2000 parts of Herba Hyperici Japonici, 250 parts of baicalins.

Crude drug in the aforementioned pharmaceutical compositions can singly be carried or mix to obtain through refining and obtain extract fully with The suitable solvent and method, and total extract is made arbitrary preparation with baicalin and mixing acceptable accessories again.The index components of gained total extract is Radix Isatidis total nitrogen, Herba Hyperici Japonici total flavones.

Radix Isatidis mentioned above can obtain Radix Isatidis extract through extracting processing with The suitable solvent, wherein solvent preferred water or ethanol, and extracting method can be infusion process, percolation, decocting method, reflux extraction or continuous extraction.Obtain Radix Isatidis extract fully as obtaining through refining by water.

The invention provides a kind of preferred for preparation technology of Radix Isatidis, specific as follows:

Get the Radix Isatidis medical material, be ground into coarse powder, decoct with water secondary, 2 hours for the first time, 1 hour for the second time, collecting decoction filtered, and filtrate is concentrated into relative density 1.15～1.20 concentrated solutions, add 1% hydrochloric acid adjust pH to 5～6, be added on the strong acid cation exchange resin column, add 2 times of water gagings towards post, discard flushing liquor, add 1% ammonia solution again, collect eluent towards post, add 1% hydrochloric acid adjust pH to 7～7.5 again, standing over night filters, concentrate, drying, promptly.The Radix Isatidis extract yield that makes by this technology is 0.5～2%, and content of total nitrogen is not less than 10% in the extract.

Radix Isatidis extract also can be by following prepared, but is not limited only to following method:

Technology one: get the Radix Isatidis medical material, be ground into coarse powder, decoct with water secondary, 2 hours for the first time, 1 hour for the second time, collecting decoction, filter, filtrate is concentrated into relative density 1.15～1.20 concentrated solutions, puts cold, add ethanol and make and contain alcohol amount and reach 80%, stir evenly, left standstill 24 hours, filter, filtrate recycling ethanol also is concentrated into relative density 1.32～1.35 thick pastes, vacuum drying, promptly.The Radix Isatidis extract yield that makes by this technology is 10～12%, and content of total nitrogen is not less than 2% in the extract.

Technology two: get the Radix Isatidis medical material, decoct with water secondary, added 12 times of amounts of water for the first time in 2 hours, added 10 times of amounts of water in 1 hour for the second time, collecting decoction filters, being evaporated to relative density is 1.03～1.11, and adding ethanol makes and contains the alcohol amount to 70%, stirs evenly, left standstill 24 hours, and filtered, filtrate recycling ethanol is to there not being the alcohol flavor, it is 8.0～8.5 that filtrate is regulated pH value with ammonia solution, stir evenly, cold preservation 48 hours adds heat extraction ammonia, it is an amount of to add water, cold preservation filters, and it is 7.0～7.5 that filtrate is regulated pH value with 1% sodium hydroxide, cold preservation filters, filtrate decompression is concentrated into the thick paste shape, spray drying, promptly.Radix Isatidis extract yield by this prepared is 2～4%, and content of total nitrogen is not less than 5% in the extract.

Herba Hyperici Japonici mentioned above can obtain Herba Hyperici Japonici extract through extracting processing with The suitable solvent, wherein solvent preferred water or ethanol, and the extracting method of Herba Hyperici Japonici can be infusion process, percolation, decocting method, reflux extraction or continuous extraction.Obtain Herba Hyperici Japonici extract fully as obtaining through refining by water.

The invention provides a kind of preferred for preparation technology of Herba Hyperici Japonici, specific as follows:

Get Herba Hyperici Japonici, cut off, decoct with water secondary, each 1 hour, collecting decoction filtered, filtrate decompression is concentrated into relative density 1.10～1.15, adds ethanol and reaches 60% to containing the alcohol amount, leaves standstill 24 hours, filter, decompression filtrate recycling ethanol is not to there being the alcohol flavor, and thin up stirs evenly to an amount of, the cold preservation standing over night, filter, filtrate is added on the polyamide column of having handled well, first water elution with 3 times of column volumes, discard eluent, 80% ethanol elution of 4 times of column volumes of reuse is collected eluent, and decompression recycling ethanol also is concentrated into the concentrated solution of relative density 1.10～1.12, spray drying, promptly.Herba Hyperici Japonici extract yield by this prepared is 1～2%, and general flavone content is not less than 60% in rutin, is not less than 20% in Quercetin.

Herba Hyperici Japonici extract also can be by following prepared, but is not limited only to following method:

Technology one: get Herba Hyperici Japonici, cut off, decoct with water secondary, each 1 hour, collecting decoction filtered, filtrate decompression is concentrated into relative density 1.10～1.15, adds ethanol and reaches 60% to containing the alcohol amount, leaves standstill 24 hours, filter, decompression filtrate recycling ethanol also is concentrated in right amount, adds ethanol and reaches 75% to containing the alcohol amount, the same method adds water to 500ml after handling, and stirs evenly, left standstill 24 hours, and filtered, filtrate adds the gelatin solution of new system, the limit edged stirs, and cold preservation 24 hours filters, filtrate is concentrated in right amount, adds ethanol and reaches 85% to containing the alcohol amount, leaves standstill 48 hours, filter, decompression filtrate recycling ethanol also is concentrated into the thick paste of relative density 1.30～1.35, vacuum drying, promptly.Herba Hyperici Japonici extract yield by this prepared is 3～5%, and general flavone content is not less than 30% in rutin, is not less than 5% in Quercetin.

Technology two: get Herba Hyperici Japonici, cut off, add 70% alcohol reflux secondary, each 2 hours, merge extractive liquid, filtered, decompression filtrate recycling ethanol also is condensed into every 1ml and is equivalent to 1g raw medicinal herbs solution, and the cold preservation standing over night filters, filtrate adds 20% sodium hydroxide adjust pH to 8.0, leaves standstill, and filters, add dilute hydrochloric acid adjust pH to 7.0 again, leave standstill, filter, filtrate decompression is concentrated into the thick paste of relative density 1.30～1.35, vacuum drying, promptly.Herba Hyperici Japonici extract yield by this prepared is 6～8%, and general flavone content is not less than 20% in rutin, is not less than 3% in Quercetin.

Pharmaceutical composition of the present invention except that available above-mentioned medical material feeds intake make, can also feed intake by the extract of the extract of Radix Isatidis and Herba Hyperici Japonici and baicalin and make.(the Radix Isatidis extract yield is 0.5～2% with respect to the yield of medical material according to extract, the Herba Hyperici Japonici extract yield is 1～2%) calculate, the present composition is by ratio of weight and the number of copies: 1～400 part of Radix Isatidis extract, 5～160 parts of Herba Hyperici Japonici extract, 60～1000 parts of baicalins; Be preferably: 3～300 parts of Radix Isatidis extracts, 10～80 parts of Herba Hyperici Japonici extract, 125～500 parts of baicalins; Optimum is: 5～140 parts of Radix Isatidis extracts, 20～40 parts of Herba Hyperici Japonici extract, 250 parts of baicalins.Radix Isatidis extract contains total nitrogen and is not less than 2%, preferably is not less than 10%; Herba Hyperici Japonici extract contains total flavones and is not less than 20% in rutin, preferably is not less than 60%, is not less than 3% in Quercetin, preferably is not less than 20%.

More than form to be by weight as proportioning, when producing, can or reduce according to the corresponding proportion increase, as large-scale production can be unit with the kilogram, or be unit with the ton, small-scale production can be unit with the gram also, weight can increase or reduce, but the constant rate of weight proportion between each composition.

More than form,, can make the preparation of 100～10000 consumptions,, can be made into 100～10000, take 1～10 at every turn,, can be made into 100～10000,1～10 of each consumption as injection as tablet as if being unit with the gram.

The ratio of above weight proportion obtains through science screening, and for especial patient, the ratio of can corresponding adjustment forming increases or reduce being no more than 100%.

The consumption of drug component of the present invention is that the inventor draws through groping in a large number to sum up, and each amounts of components all has better curative effect in above-mentioned weight portion scope.

Pharmaceutical composition of the present invention can be used for preparing the medicine for the treatment of hepatitis disease.Pharmacological testing shows, the Hepar Mus fibrosis effect of the pharmaceutical composition of the present invention Chinese People's Anti-Japanese Military and Political College is better, significantly recovers the activity of hyaluronic acid (HA) and laminin (LN); To carbon tetrachloride (CCl ₄) liver, the spleen index of poisoning mice have good reduction effect; The liver of Abensanil poisoning mice has tangible restitution; Can improve the GSH content and reduction SGPT, SGST activity of the liver of acetaminophen (AP) poisoning induced mice; There is obvious inhibition duck to infect the effect of DHBV virus.In each experiment, present composition experimental group is compared with Radix Isatidis or its extract, Herba Hyperici Japonici or its extract, baicalin group with single, enhancing evident in efficacy shows that Radix Isatidis or its extract, Herba Hyperici Japonici or its extract and baicalin three medical instruments have the effect of collaborative anti-hepatitis disease.

Pharmaceutical composition of the present invention can be mixed and made into clinically any or pharmaceutically acceptable dosage form with one or more pharmaceutically acceptable carriers, preferred oral preparation or injection are applied to the patient who needs this treatment in the mode of oral or parenteral.

When being used for oral administration, 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 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 pharmaceutical composition of the present invention is made 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.

When making injection, optional use solvent or non-aqueous solvent can add suitable additives according to the character of medicine.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.Additives commonly used comprise osmotic pressure regulator, pH value regulator, solubilizing agent, filler, antioxidant, antibacterial, emulsifying agent, suspending agent etc.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.

The present composition has the following advantages:

(1) provides a kind of pharmaceutical composition that is used for the treatment of hepatitis disease and its production and use, satisfied urgent clinical needs;

(2) first the interaction and the composition of prescription of the present composition carried out pharmacodynamic study, found that compositions has tangible antihepatitic activity, Serum ALT (glutamate pyruvate transaminase) to the liver of carbon tetrachloride poisoning induced mice), AST (glutamic oxaloacetic transaminase, GOT) has tangible restitution, filters out the weight proportion scope with significant curative effect;

(3) the present invention shows by pharmacological experiment study, and the present composition has remarkable anti-carbon tetrachloride (CCl ₄) inductive rat liver fibrosis effect, to CCl ₄Inductive chmice acute hepatic injury, Abensanil cause the chmice acute hepatic injury and have remarkable protective effect, have obvious inhibition duck to infect the effect of DHBV virus; In above-mentioned each experimental group, present composition experimental group is compared with Radix Isatidis or its extract, Herba Hyperici Japonici or its extract, baicalin group with single, enhancing evident in efficacy, it is remarkable to show that Radix Isatidis or its extract, Herba Hyperici Japonici or its extract and baicalin three medicines share anti-hepatitis effect, and consequently those skilled in the art institute is beyond thought;

(4) Radix Isatidis has the effect of heat-clearing and toxic substances removing, can antiinflammatory, enhancing immunity has therapeutical effect to hepatitis, Herba Hyperici Japonici belongs to traditional clearing away heat-damp and promoting diuresis, the subduing swelling and detoxicating medicine, have antibacterial, effect such as protect the liver; Baicalin is the anti-bacteria and anti-virus medicine, and hepatitis virus is had good inhibitory effect, three medicine drug combination determined curative effects, and reduced relative dosage, be with a wide range of applications.

Below routine by experiment beneficial effect of further setting forth medicine of the present invention.In the following experimental example: the compositions of Radix Isatidis extract, Herba Hyperici Japonici extract and baicalin is hereinafter to be referred as the BTH compositions.Used Radix Isatidis extract is taken from embodiment 1 in the experimental example, and Herba Hyperici Japonici extract is taken from embodiment 2, and baicalin is commercial, Qingan County, Heilungkiang Pharmacy stock Co., Ltd.

Experimental example 1 BTH compositions drug combination drug efficacy study---to the protective effect of rat acute hepatic injury

Animal subject: SD rat, male and female half and half, body weight 200 ± 20g.

Test sample:

The normal control group: normal saline, commercial;

CCl ₄Modeling group: CCl ₄Injection, self-control;

Diammonium glycyrrhizinate capsule group: diammonium glycyrrhizinate capsule, Jiangsu Zhengda Tianqing Drug Industry Co., Ltd

Radix Isatidis group: Radix Isatidis extract capsule, self-control;

Herba Hyperici Japonici group: Herba Hyperici Japonici extract capsule, self-control;

Baicalin group: baicalin capsule, self-control (raw material is commercial);

BTH compositions group: BTH composition capsule, self-control.

Experimental technique:

CCl ₄After entering body lipid peroxidation taking place in hepatomicrosome mainly, cause the infringement of liver plasma membrane 26S Proteasome Structure and Function, cell interior ALT, AST are overflowed, causes the active rising of ALT in the blood, AST.The hepatic injury zone is big more, and ALT, AST activity are high more.

Get 200 of SD rats, male and female half and half, precuring is 2 days under the experiment condition, is divided into 20 groups at random by sex and body weight, is respectively the normal control group, CCl ₄The modeling group, the diammonium glycyrrhizinate capsule group, the Radix Isatidis group, the Herba Hyperici Japonici group, the baicalin group, BTH compositions group: 14 groups, Radix Isatidis+Herba Hyperici Japonici+baicalin: 200mg+8000mg+1000mg, 200mg+8000mg+60mg, 500mg+4000mg+125mg, 500mg+4000mg+500mg, 1000mg+2000mg+125mg, 1000mg+2000mg+250mg, 1000mg+2000mg+500mg, 2000mg+1000mg+125mg, 2000mg+1000mg+500mg, 4000mg+500mg+1000mg, 4000mg+500mg+60mg.Each administration group is with normal saline and is configured to gastric infusion behind the suspension.Normal control group and CCl ₄The modeling group is given 0.9% normal saline, and the diammonium glycyrrhizinate group is given diammonium glycyrrhizinate capsule.Once a day, successive administration is 7 days.1h after the last administration, the normal control group is according to the dosage lumbar injection vegetable oil of 0.6ml/kg, and all the other each groups are all according to the CCl of the dosage lumbar injection 30% of 0.6ml/kg ₄Vegetable oil causes the rat acute hepatic injury.After 24 hours, eye socket is got blood, and separation of serum is measured Serum ALT (glutamate pyruvate transaminase), AST (glutamic oxaloacetic transaminase, GOT) with reitman-frankel method,

All data are handled with the SPSS10.0 statistical software.

Experimental result and conclusion: experimental result sees Table 1.

(1) normal control group and CCl ₄The modeling group is compared, and the activity of Serum ALT (glutamate pyruvate transaminase), AST (glutamic oxaloacetic transaminase, GOT) has significant difference (p＜0.01), and the modeling success is described.

(2) and CCl ₄The modeling group is compared, and the ALT of BTH compositions group, Herba Hyperici Japonici group, diammonium glycyrrhizinate capsule group, AST activity extremely significantly reduce (p＜0.01), illustrates that the hepatic injury zone extremely significantly reduces; Active significantly reduce (p＜0.05) of the ALT of Radix Isatidis group and baicalin group, AST.

(3) compare with Radix Isatidis group, Herba Hyperici Japonici group or baicalin group with single, the activity of BTH compositions group ALT, AST reduces, especially during Radix Isatidis+Herba Hyperici Japonici+baicalin=(500～15000)+(1000～4000)+(125～500), the active of ALT, AST significantly reduces, when the wherein wt proportioning is Radix Isatidis+Herba Hyperici Japonici+baicalin=(1000～7000)+2000+125, best results.

The above results shows, Radix Isatidis, Herba Hyperici Japonici and baicalin three flavor medicine compatibilities, synergistic function is arranged, and action effect is relevant with proportioning, in each parts by weight, during Radix Isatidis+Herba Hyperici Japonici+baicalin=(500～15000)+(1000～4000)+(125～500) proportioning, effect is more excellent, and when parts by weight were Radix Isatidis+Herba Hyperici Japonici+baicalin=(1000～7000)+2000+25, effect was best, points out it may be best proportioning.

Table 1 BTH composition capsule is to CCl ₄Due to Rats with Acute Liver Injury Serum ALT, the active influence of AST (X ± SD, n=10)

Group	Weight proportion (mg)	Dosage (mg/kg)	ALT(IU/L)	AST(IU/L)
					Normal control group CCl 4Modeling group diammonium glycyrrhizinate group Radix Isatidis group Herba Hyperici Japonici group baicalin group	- - - - - -	- - 15 40 40 40	24.42±2.85 ** 2800.54±314.26 1018.47±208.36 ** 2154.46±347.51 * 1208.69±123.46 ** 1987.96±355.66 *	401.77±30.12 ** 5059.38±452.74 2190.45±210.77 ** 2531.79±245.84 * 2037.85±185.63 ** 2374.56±263.15 *
BTH compositions group (Radix Isatidis+Herba Hyperici Japonici+baicalin)	200+8000+1000 200+8000+60 500+4000+125 500+4000+500 1000+2000+125 1000+2000+250 1000+2000+500 7000+2000+125 7000+2000+250 7000+2000+500 15000+1000+125 15000+1000+500 20000+500+1000 20000+500+60	40 40 40 40 40 40 40 40 40 40 40 40 40 40	1012.33±208.65 ** 812.25±119.36 ** 650.17±108.69 **&#△ 582.59±134.27 **&#△ 556.89±112.42 **&#△ 576.69±167.54 **&#△ 719.57±156.48 **＆#Δ 756.32±117.54 **&#△ 797.26±132.71 **&#△ 836.55±124.57 **&#△ 859.15±110.54 **&#△ 896.19±169.65 **&#△ 932.36±212.41 ** 1017.18±187.15 **	2174.32±187.5 ** 1832.18±163.74 ** 1792.48±143.25 **&#△ 1728.54±120.68 **&#△ 1669.78±126.59 **&#△ 1696.26±132.51 **&#△ 1716.48±149.68 **&#△ 1752.33±125.48 **&#△ 1778.94±116.15 **&#△ 1792.45±111.25 **&#△ 1847.57±184.12 **&#△ 1867.89±145.37 **&#△ 1967.89±128.46 ** 2045.47±149.25 **

Annotate: with CCl ₄The modeling group is compared, ^*P＜0.05, ^*P＜0.01; Compare with the Radix Isatidis group, ^﹠amp;P＜0.01; Compare with the Herba Hyperici Japonici group, ^#P＜0.05; ^△P＜0.05 is compared with the baicalin group.

Experimental example 2 BTH compositionss are to carbon tetrachloride (CCl ₄) influence of poisoning mice ALT, LTG, MDA

Animal subject: healthy Kunming mouse, body weight 18～22g.

Test sample:

The blank group: 0.9% normal saline, commercial;

CCl ₄Matched group: CCl ₄Injection, self-control;

Radix Isatidis extract group: Radix Isatidis extract tablet, self-control;

Herba Hyperici Japonici extract group: Herba Hyperici Japonici extract tablet, self-control;

Baicalin group: baicalin tablet, self-control (raw material is commercial);

BTH compositions group: BTH composition tablet, self-control.

Experimental technique:

Get 170 of mices, be divided into 17 groups at random, be respectively blank group, CCl ₄(12 groups of matched groups, Radix Isatidis extract group, Herba Hyperici Japonici extract group, baicalin group, BTH composite injection group, Radix Isatidis extract+Herba Hyperici Japonici extract+baicalin, 5mg+5mg+1000mg, 5mg+10mg+500mg, 5mg+20mg+250mg, 5mg+40mg+250mg, 5mg+80mg+125mg, 5mg+160mg+60mg, 140mg+5mg+1000mg, 140mg+10mg+500mg, 140mg+20mg+250mg, 140mg+40mg+250mg, 140mg+80mg+125mg, 140mg+160mg+60mg), 10 every group.Each administration group is with normal saline and is configured to gastric infusion behind the suspension.The blank group is irritated stomach 0.9% normal saline, 10ml/kg, the equal lumbar injection CCl of all the other each treated animals ₄10ml/kg, overnight fasting.Administration group gastric infusion, every day 1 time, continuous 10 days.After last administration, the sacrificed by decapitation animal is got the blood regulating liver-QI and measures serum glutamic pyruvic transminase (ALT), glyceric acid three ester (LTG), malonaldehyde (MDA) respectively.

Table 2 BTH composition tablet is to carbon tetrachloride (CCl ₄) poisoning mice ALT, LTG, MDA influence (X+SD, n=10)

Group	Parts by weight	Dosage (mg/kg)	ALT(u/dl)	LTG(mg/g)	MDA (n mol/g)
						Blank group CCl 4Matched group Radix Isatidis extract group Herba Hyperici Japonici extract group baicalin group	- - - - -	- 10 50 50 50	240±52 ** 610±247 525±210 * 455±195 * 489±202 *	15±6 ** 36±14 29±10 * 25±8 * 28±8 *	46±10 **84±26 62±15 *55±13 *59±14 *
BTH compositions group (Radix Isatidis extract+Herba Hyperici Japonici extract+baicalin)	5+5+1000 5+10+500 5+20+250 5+40+250 5+80+125 5+160+60 140+5+1000 140+10+500 140+20+250 140+40+250 140+80+125 140+160+60	50 50 50 50 50 50 50 50 50 50 50 50	301±95 **&#△ 278±82 **&#△ 250±62 **&#△ 253±64 **&#△ 283±79 **&#△ 295±87 **&#△ 288±92 **&#△ 271±69 **&#△ 254±54 **&#△ 257±50 **&#△ 285±59 **&#△ 299±89 **&#△	19±4 **&#△ 17±6 **&#△ 14±3 **&#△ 13±6 **&#△ 16±6 **&#△ 19±7 **&#△ 18±8 **&#△ 16±9 **&#△ 14±5 **&#△ 15±3 **&#△ 17±8 **&#△ 18±4 ***#△	45±13 **&#△43±12 **&#△36±8 **&#△38±5 **&#△42±10 **&#△46±15 **&#△48±11 **&#△45±10 **&#△40±6 **&#△41±4 **&#△46±7 **&#△50±8 **&#△

Annotate: with CCl ₄Matched group is compared, ^*P＜0.05, ^*P＜0.01; Compare with the Radix Isatidis extract group, ^﹠amp;P＜0.05; Compare with the Herba Hyperici Japonici extract group, ^#P＜0.05; Compare with the baicalin group, ^△P＜0.05.

Experimental result and conclusion: experimental result sees Table 2.

(1) blank group and CCl ₄The matched group utmost point significant difference (p＜0.01) of having compared illustrates the modeling success.

(2) each administration group is all to carbon tetrachloride (CCl ₄) ALT, LTG and the MDA of liver of poisoning induced mice have tangible restitution (p＜0.05 and p＜0.01).Wherein the effect of BTH compositions group all obviously is better than single with Radix Isatidis extract or Herba Hyperici Japonici extract or baicalin (p＜0.05).

Proof Radix Isatidis, Herba Hyperici Japonici and baicalin compatibility have tangible restitution to ALT, LTG and the MDA of the liver of carbon tetrachloride poisoning induced mice, make ALT, the LTG of liver and MDA level recover normal, and comparing with the blank group does not have notable difference.And relevant with the dosage ratio of compositions, Radix Isatidis extract+Herba Hyperici Japonici extract+baicalin in the BTH compositions=(effect is best during 5～140mg)+(20～40) mg+250mg.

Experimental example 3 BTH compositionss are to carbon tetrachloride (CCl ₄) influence of poisoning mice SGPT, LTG, MDA

Test sample:

Blank group: 0.9% normal saline solution, self-control;

CCl ₄Matched group: CCl ₄Injection, self-control;

Radix Isatidis extract group: Radix Isatidis extract injection, self-control;

Herba Hyperici Japonici extract group: Herba Hyperici Japonici extract injection, self-control;

Baicalin group: baicalin for injection liquid, self-control (raw material is commercial);

BTH composite injection group, self-control.

Animal subject: healthy Kunming mouse, body weight 18～22g, is divided into 14 groups at random by 140.

Experimental technique: get 140 of mices, be divided into 14 groups at random, be respectively blank group, CCl ₄Matched group, Radix Isatidis extract group, Herba Hyperici Japonici extract group, baicalin group, BTH composite injection group, 10 every group.Blank group lumbar injection 0.9% normal saline 10ml/kg, the equal lumbar injection CCl of all the other each treated animals ₄10ml/kg, overnight fasting.Administration group intraperitoneal injection, every day 1 time, dosage sees Table 1, continuous 10d, matched group is given the normal saline of respective volume, after last administration, the sacrificed by decapitation animal is got the blood regulating liver-QI and measures serum glutamic pyruvic transminase (SGPT), glyceric acid three ester (LTG), malonaldehyde (MDA) respectively.

Experimental result and conclusion: experimental result sees Table 3.

CCl ₄Matched group compares with the blank group that there were significant differences (p＜0.01), and the modeling success is described.Each administration group is all to carbon tetrachloride (CCl ₄) SGPT, LTG and the MDA of liver of poisoning induced mice have tangible restitution (p＜0.05, p＜0.01).Wherein the effect of BTH composite injection group all obviously is better than single with Radix Isatidis extract or Herba Hyperici Japonici extract or baicalin (p＜0.05).Proof Radix Isatidis, Herba Hyperici Japonici and baicalin compatibility have tangible restitution to SGPT, LTG and the MDA of the liver of carbon tetrachloride poisoning induced mice, make SGPT, the LTG of liver and MDA level recover normal, and comparing with the blank group does not have notable difference.Proof Radix Isatidis, Herba Hyperici Japonici and baicalin compatibility are evident in efficacy.

Table 3 BTH composite injection is to carbon tetrachloride (CCl ₄) poisoning mice SGPT, LTG, MDA influence (X ± SD, n=10)

Group	Parts by weight	Dosage (mg/kg)	SGPT(u/dl)	LTG(mg/g)	MDA (nmol/g)
						Blank group CCl 4Matched group Radix Isatidis extract group Herba Hyperici Japonici extract group baicalin group	- - - - -	- 10 40 40 40	242±50 602±251 ** 535±200 *# 465±185 *# 497±196 *#	14±7 34±15 ** 29±8 *# 30±9 *# 27±7 *#	45±11 83±25 **60±15 *#58±16 *#57±15 *#
BTH composite injection group (Radix Isatidis extract+Herba Hyperici Japonici extract+baicalin)	5+15+125 5+20+150 10+20+200 10+30+250 10+30+300 15+40+400 15+50+450 20+50+500 20+60+500	40 40 40 40 40 40 40 40 40	258±56 ##&△○ 305±108 ##&△○ 285±83 ##&△○ 252±51 ##&△○ 276±62 ##&△○ 274±76 ##&△○ 283±82 ##&△○ 295±96 ##&△○ 266±59 ##&△○	14±6 ##&△○ 18±5 ##&△○ 16±5 ##&△○ 13±4 ##&△○ 14±4 ##&△○ 17±5 ##&△○ 18±7 ##&△○ 19±7 ##&△○ 15±8 ##&△○	41±8 ##&△○48±11 ##&△○43±9 ##&△○38±7 ##&△○42±8 ##&△○45±10 ##&△○48±12 ##&△○49±12 ##&△○40±7 ##&△○

Annotate: ^*P＜0.05, ^*P＜0.01 is compared with the blank group; ^#P＜0.05, ^##P＜0.01 is with CCl ₄Matched group is compared; ^﹠amp;P＜0.05 is compared with the Radix Isatidis extract group; ^△P＜0.05 is compared with the Herba Hyperici Japonici extract group; ^ZeroP＜0.05 is compared with the baicalin group.

The Hepar Mus fibrosis effect of the experimental example 4 BTH compositions Chinese People's Anti-Japanese Military and Political College

Laboratory animal: the Wistar rat, body weight 180～230g, male, 90.

Test sample:

The blank group: 0.9% normal saline, commercial;

Radix Isatidis extract group: Radix Isatidis extract tablet, self-control;

Herba Hyperici Japonici extract group: Herba Hyperici Japonici extract tablet, self-control;

Baicalin group: baicalin tablet, self-control (raw material is commercial);

1 group of BTH compositions: the BTH composition tablet (Radix Isatidis+Herba Hyperici Japonici+baicalin=7g+1g+0.25g), self-control.

2 groups of BTH compositionss: the BTH composition tablet (Radix Isatidis+Herba Hyperici Japonici+baicalin=7g+2g+0.25g), self-control.

3 groups of BTH compositionss: the BTH composition tablet (Radix Isatidis+Herba Hyperici Japonici+baicalin=7g+4g+0.25g), self-control.

Experimental technique:

Get 10 of Wistar rats, subcutaneous injection Oleum Arachidis hypogaeae semen 3ml/kg, every day 1 time, totally 10 weeks, group in contrast.The Liver Fibrosis Model group is used 40%CCl ₄Peanut oil solution is pressed the 3ml/kg subcutaneous injection, 2 times weekly, in totally 8 weeks, induces Liver Fibrosis Model.Get 80 of Liver Fibrosis Model rats, be divided into 8 groups at random, be respectively model group, tiopronin group, Radix Isatidis extract group, Herba Hyperici Japonici extract group, baicalin group, 1 group of BTH compositions, 2 groups of BTH compositionss, 3 groups of BTH compositionss, 10 every group.Except that matched group, all the other each groups continue with 40%CCl ₄Peanut oil solution 3ml/kg subcutaneous injection, every day 1 time.Radix Isatidis extract group, Herba Hyperici Japonici extract group, baicalin group, 1 group of BTH compositions, 2 groups of BTH compositionss, 3 groups of whiles of BTH compositions gastric infusion relative medicine, 2 weeks of successive administration.Slaughter rat after 10 weeks, blood 2ml gets in the sterile working, separation of serum, and-20 ℃ of preservations detect hepatic fibrosis index with radioimmunology: hyaluronic acid (HA) and laminin (LN).

Experimental result and conclusion: experimental result sees Table 4.

(1) compare with the blank group, model group rat blood serum HA, LN content have utmost point significant difference, and the modeling success is described.

(2) compare with model group, tiopronin group, Herba Hyperici Japonici extract group, 1 group of BTH compositions, 2 groups of BTH compositionss, BTH compositions 3 groups of treatments heptic fibrosis rat blood serum HA and LN water mean pole significantly are lower than model group (p＜0.01), Radix Isatidis extract group and baicalin group treatment heptic fibrosis rat blood serum HA and LN level all significantly are lower than model group (p＜0.05), point out each administration group all can improve the hepatic fibrosis serological index, have the effect of anti-hepatic fibrosis.

(3) effect of 1 group of BTH compositions, 2 groups of BTH compositionss, 3 groups of BTH compositionss all obviously is better than single with Radix Isatidis extract or Herba Hyperici Japonici extract or baicalin (p＜0.05).

The above results shows that Radix Isatidis, Herba Hyperici Japonici and baicalin have synergistic function, all can improve hepatic fibrosis, has the effect of anti-hepatic fibrosis.

Table 4 BTH compositions is to CCl ₄Due to hepatic fibrosis rats serological index influence (X ± SD, n=10)

Group	Size of animal	Dosage (mg/kg)	LN (μg/L)	HA (μg/L)
					3 groups of 2 groups of BTH compositions of blank group model group Tiopronin group isatis root extract group Herba Hyperici Japonici extract group scutelloside group 1 group of BTH composition of BTH composition	10 10 10 10 10 10 10 10 10	- - 15 40 40 40 40 40 40	44.57±14.52 118.45±28.19 ○ 51.77±11.12 ** 97.15±16.24 * 80.54±17.82 ** 96.78±19.24 * 61.87±21.96 **&#△ 55.69±15.74 **&#△ 59.18±19.48 **&#△	128.59±20.47 411.74±38.13 ○ 142.12±15.57 ** 327.16±36.88 * 286.18±35.67 ** 327.79±40.56 * 193.78±26.49 **&#△ 165.87±15.29 **&#△ 188.34±26.89 **&#△

Annotate: compare with the blank group, ^ZeroP＜0.01; Compare with model group, ^*P＜0.05, ^*P＜0.01; Compare with the Radix Isatidis extract group, ^﹠amp;P＜0.05; Compare with the Herba Hyperici Japonici extract group, ^#P＜0.05; Compare with the baicalin group, ^△P＜0.05.

The influence of experimental example 5 BTH compositions Abensanil (AP) poisoning mice SGPT, SGST activity and GSH content

Animal subject: healthy Kunming mouse, body weight 18～22g.

Test sample:

Blank group: 0.9% normal saline solution, self-control;

AP matched group: AP injection, self-control;

Radix Isatidis extract group: Radix Isatidis extract injection, self-control;

Herba Hyperici Japonici extract group: Herba Hyperici Japonici extract injection, self-control;

Baicalin group: baicalin for injection liquid, self-control (raw material is commercial);

BTH composite injection group (Radix Isatidis extract+Herba Hyperici Japonici extract+baicalin=10mg+30mg+250mg): be divided into basic, normal, high three dosage groups, self-control.

Animal subject: healthy Kunming mouse, body weight 18～22g, is divided into 8 groups at random by 80.

Experimental technique:

Get 80 of mices, be divided into 8 groups at random, be respectively blank group, AP matched group, Radix Isatidis extract group, Herba Hyperici Japonici extract group, baicalin group, basic, normal, high three the dosage groups of BTH composite injection, 10 every group.Except that the blank group, the equal lumbar injection AP100ml/kg of all the other each treated animals, overnight fasting.Administration group intraperitoneal injection, every day 1 time, dosage sees Table 3, continuous 10d, matched group is given the normal saline of respective volume, after last administration, lumbar injection AP100mg/kg, behind the fasting 3h, the sacrificed by decapitation animal is got the blood regulating liver-QI and measures serum glutamic pyruvic transminase (SGPT), glutathione S-transferase (SGST) and reduced glutathion (GSH) respectively.

The influence of table 5 BTH compositions Abensanil (AP) poisoning mice SGPT, SGST activity and GSH content (X ± SD, n=10)

Group	Dosage (mg/kg)	SGPT (u/dl)	SGST (nmol·min·ml -1)	GSH (mmol/g liver)
					Dosage group BTH composition high dose group in the blank group AP control group isatis root extract group Herba Hyperici Japonici extract group scutelloside group BTH composition low dose group BTH composition	- 100 50 50 50 30 50 80	176±30 724±312 **435±214 *#444±231 *#461±239 *#289±165 ##&△○252±160 ##&△○219±157 ##&△○	116±28 514±253 ** 305±163 *# 284±174 *# 297±159 *# 175±88 ##&△○ 158±74 ##&△○ 142±70 ##&△○	4.6±1.6 2.5±1.1 **3.3±1.2 *#3.1±1.3 *#2.9±1.0 *#4.3±1.6 ##&△○4.8±1.5 ##&△○5.3±1.7 ##&△○

Annotate: ^*P＜0.05, ^*P＜0.01 is compared with the blank group; ^#P＜0.05, ^##P＜0.01 is compared with the AP matched group; ^﹠amp;P＜0.05 is compared with the Radix Isatidis extract group; ^△P＜0.05 is compared with the Herba Hyperici Japonici extract group; ^ZeroP＜0.05 is compared with the baicalin group.

Experimental result and conclusion: experimental result sees Table 5.

(1) compare with the blank group, the AP matched group extremely significantly reduces GSH content and the utmost point significantly improves SGPT, SGST activity (p＜0.01), and the modeling success is described.

(2) compare with the blank group, each administration group can both improve the GSH content and reduction SGPT, SGST activity (p＜0.05, p＜0.01) of the liver of acetaminophen (AP) poisoning induced mice.

(3) wherein the effect of BTH composite injection group all obviously is better than single with Radix Isatidis extract or Herba Hyperici Japonici extract or baicalin (p＜0.05).The liver of proof Radix Isatidis, Herba Hyperici Japonici and baicalin compatibility Abensanil poisoning mice has tangible restitution, makes SGPT, the SGST level of liver recover normal, and comparing with the blank group does not have notable difference.Along with the increase of dosage, increase GSH content and reduce the active effect increasing of SGPT, SGST, effect is best during high dose.

Experimental example 6 BTH compositionss are to carbon tetrachloride (CCl ₄) liver, the shadow of spleen index of acute liver damage mice due to the poisoning Ring

Animal subject: healthy Kunming mouse, body weight 18～22g.

Test sample:

Blank group: 0.9% normal saline solution, self-control;

CCl ₄Matched group: CCl ₄Injection, self-control;

Radix Isatidis extract group: Radix Isatidis extract injection, self-control;

Herba Hyperici Japonici extract group: Herba Hyperici Japonici extract injection, self-control;

Baicalin group: baicalin for injection liquid, self-control (raw material is commercial);

BTH composite injection group (Radix Isatidis extract+Herba Hyperici Japonici extract+baicalin=10mg+30mg+250mg): be divided into basic, normal, high three dosage groups, self-control.

Animal subject: healthy Kunming mouse, body weight 18～22g, is divided into 8 groups at random by 80.

Experimental technique:

Get 80 of mices, be divided into 8 groups at random, be respectively blank group, CCl ₄Matched group, Radix Isatidis extract group, Herba Hyperici Japonici extract group, baicalin group, basic, normal, high three the dosage groups of BTH composite injection, 10 every group.Blank group lumbar injection 0.9% normal saline 10ml/kg, the equal lumbar injection CCl of all the other each treated animals ₄10ml/kg, overnight fasting.Administration group intraperitoneal injection, every day 1 time, dosage sees Table 2, continuous 10d, matched group is given the normal saline of respective volume, and after last administration, the sacrificed by decapitation animal is got liver and spleen, inhales the liquid of dehematizing, and cuts off fat, mesentery, accurately weighs.

Table 6 BTH compositions is to carbon tetrachloride (CCl ₄) influence of liver, the spleen index of acute liver damage mice due to the poisoning (X ± SD, n=10)

Group	Number of animals (only)	Dosage (mg/kg)	Liver index (g/100g)	Spleen index (g/100g)
					Blank group CCl 4Dosage group BTH composition high dose group in the control group isatis root extract group Herba Hyperici Japonici extract group scutelloside group BTH composition low dose group BTH composition	10 10 10 10 10 10 10 10	- 10 50 50 50 30 50 80	6.15±1.00 8.45±0.65 **7.52±0.72 *#7.29±0.70 *#7.38±0.75 *#6.65±0.80 ##&△○6.46±0.78 ##&△○6.32±0.74 ##&△○	0.630±0.142 0.742±0.076 **0.695±0.043 *#0.701±0.052 *#0.706±0.048 *#0.654±0.056 ##&△○0.641±0.046 ##&△○0.631±0.055 ##&△○

Annotate: ^*P＜0.05, ^*P＜0.01 is compared with the blank group; ^#P＜0.05, ^##P＜0.01 is with CCl ₄Matched group is compared; ^﹠amp;P＜0.05 is compared with the Radix Isatidis extract group; ^△P＜0.05 is compared with the Herba Hyperici Japonici extract group; ^ZeroP＜0.05 is compared with the baicalin group.

Experimental result and conclusion: experimental result sees Table 6.

(1) compares CCl with the blank group ₄Matched group liver, the spleen index utmost point significantly improve (p＜0.01), and the modeling success is described.

(2) and CCl ₄Matched group is compared, and each administration group can both significantly reduce carbon tetrachloride (CCl ₄) liver, the spleen index (p＜0.05 and p＜0.01) of poisoning mice.Wherein the effect of BTH composite injection group all obviously is better than single with Radix Isatidis extract or or Herba Hyperici Japonici extract or baicalin (p＜0.05).

Proof Radix Isatidis, Herba Hyperici Japonici and baicalin compatibility have good reduction carbon tetrachloride (CCl ₄) liver of poisoning mice, the effect of spleen index, comparing with the blank group does not have significant difference.And relevant with the dosage of compositions, effect is best during high dose.

Experimental example 7 BTH compositionss are to the inhibitory action of DHBV-DNA

Animal subject: commercially available 1 age in days Beijing duck.

Test sample:

Blank group: 0.9% normal saline solution, self-control;

Positive controls: acyclovir (ACV) injection, self-control;

Radix Isatidis extract group: Radix Isatidis extract injection, self-control;

Herba Hyperici Japonici extract group: Herba Hyperici Japonici extract injection, self-control;

Baicalin group: baicalin for injection liquid, self-control (raw material is commercial);

BTH composite injection group (Radix Isatidis extract+Herba Hyperici Japonici extract+baicalin=10mg+30mg+250mg): be divided into basic, normal, high three dosage groups, self-control.

Dosage unit: mg/kg.

Virus: DHBV positive serum.

Experimental technique:

(1) animal model: Beijing duck is got blood through sufficient intravenous injection 0.2mlDHBV positive serum behind the 7d, and separation of serum is preserved check for-20 ℃.

(2) Drug therapy: filter out 42 of the positive ducks that infect successfully, be divided into 7 groups at random, 6 every group.Be respectively blank group, positive controls, Radix Isatidis extract group, Herba Hyperici Japonici extract group, baicalin group, basic, normal, high three the dosage groups of BTH composite injection, dosage sees the following form.Intravenous administration, 2 weeks of administration.Respectively at before the medicine, after medication the 7th day, medication the 14th day and the drug withdrawal the 3rd day, from duck lower limb vein haemospasia, separation of serum ,-20 ℃ of preservations are to be tested.

(3) detection method: adopt DHBV-DNA Dot Blot method, with hybridization spot absorbance (A) as specimen DHBV-DNA level value.

(4) statistical procedures: relatively, adopt paired t-test before medication group different time dna content and the medication; Same time D NA content of medication group and virus control group relatively adopt the t check.

Experimental result and conclusion: experimental result sees Table 7.

(1) compare with the blank group, each administration group all obviously suppresses DHBV virus (p＜0.05, p＜0.01).

(2) compare with Radix Isatidis extract or Herba Hyperici Japonici extract or baicalin with single, the effect of BTH composite injection group all obviously is better than single with Radix Isatidis extract or Herba Hyperici Japonici extract or baicalin (p＜0.05).(3) with on the same group compare before the treatment, do not have after the drug withdrawal before knock-on and the administration comparing difference significantly ( ^*P＜0.05 He ^*And positive controls does not relatively have significant difference after drug withdrawal with before the administration p＜0.01).

In sum, Radix Isatidis, Herba Hyperici Japonici and baicalin compatibility can suppress DHBV-DNA preferably, do not have knock-on after the drug withdrawal, and effect is better, and curative effect is relevant with the dosage of compositions, and effect of high dosage is best.

Table 7 BTH compositions to the inhibitory action of DHBV-DNA (X ± SD, n=10)

Group	Dosage	DHBV-DN A titre before the treatment	Treatment back 7d DHBV-DNA titre	Treatment back 14d DHBV-DNA titre	3d DHBV-DNA titre after the drug withdrawal
						Dosage group BTH composition high dose group in the blank group positive controls isatis root extract group Herba Hyperici Japonici extract group scutelloside group BTH composition low dose group BTH composition	- 20 50 50 50 30 50 80	1.65±0.05 1.63±0.04 1.59±0.03 1.62±0.06 1.61±0.05 1.58±0.05 1.61±0.04 1.62±0.03	1.61±0.02 0.78±0.15 **&&1.35±0.06 *&1.30±0.07 *&1.28±0.09 *&1.01±0.10 **&&#△○0.94±0.15 **&&#△○0.82±0.15 **&&#△○	1.63±0.04 0.79±0.14 **&&1.30±0.07 *&1.26±0.08 *&1.24±0.05 *&0.99±0.11 **&&#△○0.92±0.13 **&&#△○0.80±0.14 **&&#△○	1.64±0.03 1.62±0.04 1.41±0.02 *&1.35±0.06 *&1.39±0.06 *&1.05±0.08 **&&#△○0.99±0.08 **&&#△○0.92±0.10 **&&#△○

Annotate: ^*P＜0.05, ^*P＜0.01, with on the same group the treatment before compare; ^﹠amp;P＜0.05, ^{﹠amp; ﹠amp;}P＜0.01 is compared with the blank group; ^#P＜0.05 is compared with the Radix Isatidis extract group; ^△P＜0.05 is compared with the Herba Hyperici Japonici extract group; ^ZeroP＜0.05 is compared with the baicalin group.

Experimental example 8 injected in mice administration acute toxicity testings

(1) experimental technique

Test sample: the BTH composite injection (self-control, 5ml: Radix Isatidis extract+Herba Hyperici Japonici extract+baicalin=10mg+30mg+250mg).

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.

Observation item: death toll, general state, body weight, cut open inspection, median lethal dose(LD 50).

(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 maximum dosage-feeding experiment in a day.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 1,3,7,14 day 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, infer that the BTH composite injection is 0.4ml/10g to the maximum tolerated dose of male and female mouse vein and intraperitoneal injection, be 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 9 BTH composite injection stability experiments

Sample: the BTH composite injection (self-control, 5ml: Radix Isatidis extract+Herba Hyperici Japonici extract+baicalin=10mg+30mg+250mg).

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, the BTH composite injection is more stable.

4, the specific embodiment

The specific embodiment of form is described in further detail foregoing of the present invention by the following examples.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.Used Radix Isatidis extract is taken from embodiment 1 in the experimental example among the embodiment 3～10, and Herba Hyperici Japonici extract is taken from embodiment 2, and baicalin is commercial, Qingan County, Heilungkiang Pharmacy stock Co., Ltd.

The preparation of embodiment 1 Radix Isatidis extract

The preparation method of Radix Isatidis extract

Get the Radix Isatidis medical material, be ground into coarse powder, decoct with water secondary, 2 hours for the first time, 1 hour for the second time, collecting decoction filtered, and filtrate is concentrated into relative density 1.15～1.20 concentrated solutions, add 1% hydrochloric acid adjust pH to 5～6, be added on the strong acid cation exchange resin column, add 2 times of water gagings towards post, discard flushing liquor, add 1% ammonia solution again, collect eluent towards post, add 1% hydrochloric acid adjust pH to 7～7.5 again, standing over night filters, concentrate, drying, promptly.

The Radix Isatidis extract identification experiment

Get this product 0.1g, add ethanol 10ml, supersound process 15min, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Other gets Radix Isatidis control medicinal material 1g, adds ethanol 30ml and shines medical material solution in pairs with legal system.Get L-proline reference substance again, add 80% ethanol and make the solution that every 1ml contains 2mg, in contrast product solution.Drawing above-mentioned three kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, is developing solvent with n-butanol-water-glacial acetic acid (4: 1: 1), launches, and takes out, and dries, and spray is with 0.2% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of comparison film chromatograph on, show the speckle of same color respectively.

The Radix Isatidis extract assay

Content of total nitrogen is measured

Total nitrogen is measured, and measures according to the pharmacopeia N2 method to get final product.

According to above-mentioned technology, make Radix Isatidis extract three batch samples, its content and yield see Table 8.Radix Isatidis extract yield by this prepared is 0.5～2%, and wherein content of total nitrogen is not less than 10%.

Table 8 Radix Isatidis extract total nitrogen content and yield

Lot number	Total nitrogen content (%)	Yield (%)
			123 is average	15.67 16.13 16.51 16.10	1.54 1.31 1.34 1.40

The preparation of embodiment 2 Herba Hyperici Japonici extract

The preparation method of Herba Hyperici Japonici extract

Get Herba Hyperici Japonici (Herba Hyperici Japonici), cut off, decoct with water secondary, each 1 hour, collecting decoction filtered, filtrate decompression is concentrated into relative density 1.10～1.15, adds ethanol and reaches 60% to containing the alcohol amount, leaves standstill 24 hours, filter, decompression filtrate recycling ethanol is not to there being the alcohol flavor, and thin up stirs evenly to an amount of, the cold preservation standing over night, filter, filtrate is added on the polyamide column of having handled well, first water elution with 3 times of column volumes, discard eluent, 80% ethanol elution of 4 times of column volumes of reuse is collected eluent, and decompression recycling ethanol also is concentrated into the concentrated solution of relative density 1.10～1.12, spray drying, promptly.

The Herba Hyperici Japonici extract identification experiment

Get this product 10ml, add methanol 10ml, hydrochloric acid 1ml, reflux 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 5ml makes dissolving, as need testing solution.Other gets the Quercetin reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw need testing solution 5 μ l, reference substance solution 1 μ l, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-formic acid (5: 2: 1) is developing solvent, launches, and takes out, dry, spray, is put under the ultra-violet lamp (365nm) and is inspected about 5 minutes of 105 ℃ of heating with 1% aluminum chloride alcoholic solution.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

The Herba Hyperici Japonici extract assay

Measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000).

Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Methanol-0.2% phosphoric acid solution (50: 50) is a mobile phase; The detection wavelength is 260nm.Number of theoretical plate calculates by the Quercetin peak should be not less than 2500.

The preparation of reference substance solution: precision takes by weighing through 24 hours Quercetin reference substance of phosphorus pentoxide drying under reduced pressure, adds methanol and makes the solution that every 1ml contains 50 μ g, promptly.

The preparation of need testing solution: precision takes by weighing this product 0.2g, puts in the round-bottomed flask, adds methanol 15ml, hydrochloric acid 1ml, mixing, reflux 30 minutes is put cold, move in the 50ml measuring bottle, be diluted to scale, shake up with methanol, filter, precision is measured subsequent filtrate 5ml, puts in the 50ml measuring bottle, be diluted to scale with methanol, shake up, filter, promptly.

Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.

According to above-mentioned technology, make Herba Hyperici Japonici extract three batch samples, its content and yield see Table 9.Herba Hyperici Japonici extract extract yield by this prepared: 1～2%, general flavone content (in rutin): be no less than 60%; General flavone content (in Quercetin): be no less than 20%.

Table 9 Herba Hyperici Japonici extract yield and general flavone content

Lot number	Yield (%)	General flavone content (in rutin) (%)	General flavone content (in Quercetin) (%)
				123 is average	1.06 1.53 1.96 1.52	68.49 72.15 75.63 72.09	26.12 27.36 29.64 27.71

The preparation of embodiment 3 BTH composition tablets

Prescription one:

Radix Isatidis extract	14g (suitable and crude drug 1000g)
		Herba Hyperici Japonici extract	30g (suitable and crude drug 2000g)
Baicalin	125g
		Starch	110.0g
Microcrystalline Cellulose	35.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	2.0g
		Carboxymethylstach sodium	4.0g
Preparation altogether	1000

Prescription two:

Radix Isatidis extract	14g (suitable and crude drug 1000g)
		Herba Hyperici Japonici extract	30g (suitable and crude drug 2000g)
Baicalin	250g
		Starch	120.0g
Microcrystalline Cellulose	40.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	2.0g
		Carboxymethylstach sodium	4.0g
Preparation altogether	1000

Prescription three:

Radix Isatidis extract	14g (suitable and crude drug 1000g)
		Herba Hyperici Japonici extract	30g (suitable and crude drug 2000g)
Baicalin	500g
		Starch	130.0g
Microcrystalline Cellulose	50.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	3.0g
		Carboxymethylstach sodium	5.0g
Preparation altogether	1000

Prescription four:

Radix Isatidis extract	100g (suitable and crude drug 7000g)
		Herba Hyperici Japonici extract	30g (suitable and crude drug 2000g)
Baicalin	125g
		Starch	120.0g
Microcrystalline Cellulose	40.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	2.0g
		Carboxymethylstach sodium	4.0g
Preparation altogether	1000

Prescription five:

Radix Isatidis extract	100g (suitable and crude drug 7000g)
		Herba Hyperici Japonici extract	30g (suitable and crude drug 2000g)
Baicalin	250g
		Starch	130.0g
Microcrystalline Cellulose	45.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	2.0g
		Carboxymethylstach sodium	4.0g
Preparation altogether	1000

Prescription six:

Radix Isatidis extract	100g (suitable and crude drug 7000g)
		Herba Hyperici Japonici extract	30g (suitable and crude drug 2000g)
Baicalin	500g
		Starch	150.0g
Microcrystalline Cellulose	55.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	3.0g
		Carboxymethylstach sodium	6.0g
Preparation altogether	1000

Preparation technology:

1) it is standby Radix Isatidis extract, Herba Hyperici Japonici extract and baicalin to be pulverized 100 mesh sieves.

2) take by weighing raw material and adjuvant according to recipe quantity.

3) hypromellose 2% the aqueous solution made soluble in water is standby.

4) with Radix Isatidis extract, Herba Hyperici Japonici extract, baicalin, starch, microcrystalline Cellulose mix homogeneously, adding 2%HPMC aqueous solution is an amount of, stirs, and makes suitable soft material

5) 20 mesh sieve system granules.

6) grain is dried under 60 ℃ condition.

7) dry good granule adds magnesium stearate and carboxymethylstach sodium, crosses 18 mesh sieve granulate, mix homogeneously.

8) sample, the semi-finished product chemical examination.

9) the sheet weight sheet of determining according to chemical examination.

10) finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 4 BTH composition capsules

Prescription one:

Radix Isatidis extract	10g
		Herba Hyperici Japonici extract	20g
Baicalin	250g
		Starch	60.0g
Microcrystalline Cellulose	20.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	1.0g
		Preparation altogether	1000

Prescription two:

Radix Isatidis extract	70g
		Herba Hyperici Japonici extract	20g
Baicalin	250g
		Starch	65.0g
Microcrystalline Cellulose	25.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	2.0g
		Preparation altogether	1000

Prescription three:

Radix Isatidis extract	10g
		Herba Hyperici Japonici extract	30g
Baicalin	250g
		Starch	60.0g
Microcrystalline Cellulose	20.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	1.0g
		Preparation altogether	1000

Prescription four:

Radix Isatidis extract	70g
		Herba Hyperici Japonici extract	30g
Baicalin	250g
		Starch	65.0g
Microcrystalline Cellulose	25.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	2.0g
		Preparation altogether	1000

Prescription five:

Radix Isatidis extract	10g
		Herba Hyperici Japonici extract	40g
Baicalin	250g
		Starch	60.0g
Microcrystalline Cellulose	20.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	1.0g
		Preparation altogether	1000

Prescription six:

Radix Isatidis extract	70g
		Herba Hyperici Japonici extract	40g
Baicalin	250g
		Starch	65.0g
Microcrystalline Cellulose	25.0g
		The 2%HPMC aqueous solution	In right amount
Magnesium stearate	2.0g
		Preparation altogether	1000

Preparation technology:

1) it is standby Radix Isatidis extract, Herba Hyperici Japonici extract and baicalin to be pulverized 100 mesh sieves.

2) take by weighing raw material and adjuvant according to recipe quantity.

3) hypromellose 2% the aqueous solution made soluble in water is standby.

4) with Radix Isatidis extract, Herba Hyperici Japonici extract, baicalin, starch, microcrystalline Cellulose mix homogeneously, adding 2%HPMC aqueous solution is an amount of, stirs, and makes suitable soft material.

5) 20 mesh sieve system granules.

6) grain is dried under 60 ℃ condition.

7) dry good granule adds magnesium stearate, crosses 18 mesh sieve granulate, mix homogeneously.

8) sample, the semi-finished product chemical examination.

9) loading amount of determining according to chemical examination incapsulates.

10) finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 5 BTH composition granules

Prescription one:

Radix Isatidis	7000g
		Herba Hyperici Japonici	1000g
Baicalin	250g
		Icing Sugar	500.0g
Dextrin	300.0g
		Tangerine sugar	20g
The 2%HPMC50% alcoholic solution	In right amount
		Preparation altogether	1000 bags

Prescription two:

Radix Isatidis	7000g
		Herba Hyperici Japonici	2000g
Baicalin	250g
		Icing Sugar	600.0g
Dextrin	350.0g
		Tangerine sugar	25g
The 2%HPMC50% alcoholic solution	In right amount
		Preparation altogether	1000 bags

Prescription three:

Radix Isatidis	7000g
		Herba Hyperici Japonici	4000g
Baicalin	250g
		Icing Sugar	800.0g
Dextrin	400.0g
		Tangerine sugar	30g
The 2%HPMC50% alcoholic solution	In right amount
		Preparation altogether	1000 bags

Preparation technology:

1) it is standby sucrose to be pulverized 100 mesh sieves.It is standby that Radix Isatidis extract, Herba Hyperici Japonici extract and baicalin were pulverized 100 mesh sieves.

2) take by weighing raw material and adjuvant according to recipe quantity.

3) the method mix homogeneously that Radix Isatidis extract, Herba Hyperici Japonici extract, baicalin and Icing Sugar, dextrin, tangerine sugar are progressively increased with equivalent, adding 2%HPMC50% alcoholic solution is an amount of, stirs, and makes suitable soft material.

4) 20 mesh sieve system granules.

5) grain is dried under 60 ℃ condition.

6) granule is crossed 18 mesh sieve granulate.

7) sample, the content of principal agent is determined loading amount in the semi-finished product chemical examination granule.

8) packing, finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 6 BTH composition oral liquid

Prescription one:

Radix Isatidis extract	10g
		Herba Hyperici Japonici extract	30g
Baicalin	250g
		Polyoxyethylene sorbitan monoleate	50g
Sodium benzoate	15g
		Stevioside	10g
Water	Add to 1000ml
		Preparation altogether	1000ml

Prescription two:

Radix Isatidis extract	70g
		Herba Hyperici Japonici extract	30g
Baicalin	250g
		Polyoxyethylene sorbitan monoleate	55g
Sodium benzoate	20g
		Stevioside	15g
Water	Add to 1000ml
		Preparation altogether	1000ml

Prescription three:

Radix Isatidis extract	140g
		Herba Hyperici Japonici extract	30g
Baicalin	250g
		Polyoxyethylene sorbitan monoleate	70g
Sodium benzoate	35g
		Stevioside	30g
Water	Add to 1000ml
		Preparation altogether	1000ml

Preparation technology:

1) polyoxyethylene sorbitan monoleate is complete with the water dissolution of dosing amount 40%, again Radix Isatidis extract, Herba Hyperici Japonici extract are added, the heated and stirred dissolving is fully.Baicalin adds that to add an amount of dissolution of sodium hydroxide after the low amounts of water heating complete.

2) sodium formate and stevioside are complete with the water dissolution of dosing amount 20%.

3) and above-mentioned solution, mend and add water to full dose.

4) filtering with microporous membrane of 0.8 μ m.

5) finished product chemical examination.

6) fill.Finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 7 BTH compositions aqueous injection

Prescription:

Radix Isatidis extract	10g
		Herba Hyperici Japonici extract	30g
Baicalin	250g
		Polyoxyethylene sorbitan monoleate	50g
Water for injection	Add to 5000ml
		Preparation altogether	1000

Preparation technology:

1) carries 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.

2) get the water for injection of dosing amount 40%, add polyoxyethylene sorbitan monoleate, Radix Isatidis extract, the Herba Hyperici Japonici extract of recipe quantity, the heated and stirred dissolving fully.Baicalin adds a small amount of water for injection, and an amount of dissolution of sodium hydroxide in heating back merges solution, and benefit adds to the full amount of water for injection.

3) go into the needle-use activated carbon of dosing amount 0.1%, heated and stirred 15 minutes.

4) sand filtration rod filtering decarbonization.Measure the also pH value of regulator solution.

5) the microporous filter membrane fine straining of 0.45 μ m.

6) look into the clarity of solution, the semi-finished product chemical examination.

7) the solution sealing by fusing is in glass ampule.

8) 00 ℃ of flowing steam sterilization is 30 minutes.

9) heat is put into 0.01% methylene blue solution with sample and is hunted leak.

10) lamp inspection, finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 8 BTH composition powder injections

Prescription:

Radix Isatidis extract	10g
		Herba Hyperici Japonici extract	30g
Baicalin	250g
		Polyoxyethylene sorbitan monoleate	50g
Mannitol	300g
		Sterile water for injection	Add to 3000ml
Preparation altogether	1000

Preparation technology:

1) vessel of at first dosing being used and antibiotic glass bottle, plug etc. carry out aseptic process.

2) take by weighing raw material and adjuvant according to recipe quantity.

3) water for injection of getting dosing amount 40% adds polyoxyethylene sorbitan monoleate, Radix Isatidis extract, the Herba Hyperici Japonici extract of recipe quantity, and the heated and stirred dissolving fully.Baicalin adds a small amount of water for injection, an amount of dissolution of sodium hydroxide in heating back.Merge above-mentioned solution, add the dissolving of mannitol heated and stirred more fully, add sterile water for injection to full dose.

4) needle-use activated carbon of adding dosing amount 0.1%, heated and stirred 15 minutes.

5) through sand filtration rod filtering decarbonization.Measure the also pH value of regulator solution.

6) through the microporous filter membrane fine straining of 0.22 μ m.

7) clarity of inspection solution, the semi-finished product chemical examination.

8) 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.

9) lyophilizing finishes, and lid is rolled in tamponade.

10) finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 9 BTH compositions sodium chloride transfusion

Prescription:

Radix Isatidis extract	10g
		Herba Hyperici Japonici extract	30g
Baicalin	250g
		Polyoxyethylene sorbitan monoleate	50g
Sodium chloride	900g
		Water for injection	Add to 100000ml
Preparation altogether	1000 bottles

Preparation technology:

1) handles the previous day such as pipeline that dosing uses and container etc., face with the fresh water for injection flushing of preceding reuse.

2) get the water for injection of dosing amount 40%, add polyoxyethylene sorbitan monoleate, Radix Isatidis extract, the Herba Hyperici Japonici extract of recipe quantity, the heated and stirred dissolving fully.Baicalin adds a small amount of water for injection, an amount of dissolution of sodium hydroxide in heating back.Sodium chloride is complete with the water for injection dissolving of dosing amount 40%.Merge above-mentioned solution, benefit adds to the full amount of water for injection.

3) go into the needle-use activated carbon of dosing amount 0.1%, heated and stirred 15 minutes.

4) sand filtration rod filtering decarbonization.Measure the also pH value of regulator solution.

5) through the microporous filter membrane fine straining of 0.45 μ m.

6) clarity of inspection solution, the semi-finished product chemical examination.

7) fill is in the infusion bottle of 100ml.

8) 115 ℃ of pressure sterilizings are 30 minutes.

9) lamp inspection, finished product is examined entirely, the packing warehouse-in.

The preparation of embodiment 10 BTH compositions glucose infusion liquids

Prescription:

Radix Isatidis extract	10g
		Herba Hyperici Japonici extract	30g
Baicalin	250g
		Polyoxyethylene sorbitan monoleate	50g
Glucose	5000g
		Water for injection	Add to 100000ml
Preparation altogether	1000 bottles

Preparation technology:

1) carries 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.

2) get the water for injection of dosing amount 40%, add polyoxyethylene sorbitan monoleate, Radix Isatidis extract, the Herba Hyperici Japonici extract of recipe quantity, the heated and stirred dissolving fully.Baicalin adds a small amount of water for injection, an amount of dissolution of sodium hydroxide in heating back.Glucose is complete with the water for injection dissolving of dosing amount 40%.Merge above-mentioned solution, benefit adds to the full amount of water for injection.

3) needle-use activated carbon of adding dosing amount 0.1%, heated and stirred 15 minutes.

4) through sand filtration rod filtering decarbonization.Measure the also pH value of regulator solution.

5) through the microporous filter membrane fine straining of 0.45 μ m.

6) clarity of inspection solution, the semi-finished product chemical examination.

7) fill is in the infusion bottle of 100ml.

8) 115 ℃ of pressure sterilizings are 30 minutes.

9) lamp inspection, finished product is examined entirely, the packing warehouse-in.

Claims (10)

1, a kind of pharmaceutical composition of anti-hepatitis is characterized in that, the crude drug of making its contained composition and effectiveness is Radix Isatidis, Herba Hyperici Japonici and baicalin, and its parts by weight are: 200～20000 parts of Radix Isatidis, 500～8000 parts of Herba Hyperici Japonici, 60～1000 parts of baicalins.

2, pharmaceutical composition as claimed in claim 1 is characterized in that, the parts by weight of crude drug are: 500～15000 parts of Radix Isatidis, 1000～4000 parts of Herba Hyperici Japonici, 125～500 parts of baicalins.

3, pharmaceutical composition as claimed in claim 2 is characterized in that, the parts by weight of crude drug are: 1000～7000 parts of Radix Isatidis, 2000 parts of Herba Hyperici Japonici, 250 parts of baicalins.

4, as the described arbitrary preparation of drug combination method of claim 1～3, it is characterized in that, described Radix Isatidis and Herba Hyperici Japonici can be with The suitable solvent respectively or mix through extracting processing and obtain its extract, total extract is made preparation with baicalin and pharmaceutic adjuvant hybrid process again, and the index components of gained total extract is Radix Isatidis total nitrogen, Herba Hyperici Japonici total flavones.

5, pharmaceutical composition as claimed in claim 1, it is characterized in that, this pharmaceutical composition can also be made by Radix Isatidis extract, Herba Hyperici Japonici extract and baicalin, and its parts by weight are: 1～400 part of Radix Isatidis extract, 5～160 parts of Herba Hyperici Japonici extract, 60～1000 parts of baicalins.

6, pharmaceutical composition as claimed in claim 6 is characterized in that, its parts by weight are: 3～300 parts of Radix Isatidis extracts, 10～80 parts of Herba Hyperici Japonici extract, 125～500 parts of baicalins.

7, pharmaceutical composition as claimed in claim 7 is characterized in that, its parts by weight are: 5～140 parts of Radix Isatidis extracts, 20～40 parts of Herba Hyperici Japonici extract, 250 parts of baicalins.

As the described arbitrary pharmaceutical composition of claim 5～7, it is characterized in that 8, Radix Isatidis extract contains the Radix Isatidis total nitrogen and is not less than 2%; Herba Hyperici Japonici extract contains general flavone content and is not less than 20% in rutin, is not less than 3% in Quercetin.

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

10, pharmaceutical composition according to claim 9 is characterized in that, clinically described or pharmaceutically acceptable dosage form is oral formulations or injection.