CN101002849B - Traditional Chinese medicine extractive or tablet for treating hepatitis, and its preparing method - Google Patents
Traditional Chinese medicine extractive or tablet for treating hepatitis, and its preparing method Download PDFInfo
- Publication number
- CN101002849B CN101002849B CN2007100029796A CN200710002979A CN101002849B CN 101002849 B CN101002849 B CN 101002849B CN 2007100029796 A CN2007100029796 A CN 2007100029796A CN 200710002979 A CN200710002979 A CN 200710002979A CN 101002849 B CN101002849 B CN 101002849B
- Authority
- CN
- China
- Prior art keywords
- radix
- filtrate
- effective site
- fructus gardeniae
- herba artemisiae
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Abstract
A Chinese medicine in the form of tablet for treating hepatitis hepatocirrhosis, biliary tract infection and cholelithiasis is prepared from 6 Chinese-medicinal materials including capejasmine fruit, scutellaria root, rhubard, coptis root, etc. Its preparing process including extracting their active components is also disclosed.
Description
Technical field:
The present invention relates to a kind of Chinese medicine extract for the treatment of hepatitis, tablet and preparation method thereof technical field, belong to technical field of Chinese medicines.
Background technology:
Hepatitis is common clinical, frequently-occurring disease.China is world hepatitis big country, and global hepatitis 80% concentrates on China.According to seroepidemiological survey, it is 80.9% that hepatitis A virus infects prevalence rate, and hepatitis b virus infected prevalence rate reaches 57.6%, and hepatitis B surface antigen (HBsAg) positive rate is 9.75%, about 1.2 hundred million people.The infection with hepatitis C virus rate is 32%, and fourth type and hepatitis E also have popular.Because hepatitis particularly chronic hepatitis route of transmission complexity, the course of disease is long, is easy to repeatedly and chronicity, and is closely related with liver cirrhosis and hepatocarcinoma.Annual because of dead about 300,000 people of hepatopathy.Therefore, the hepatitis particularly control of viral hepatitis is the magnificent mission that China infectious disease worker bears always, also is the difficult point and the focus of new drug research exploitation always.
Summary of the invention:
For better service in hepatitis, overcome the inconvenience of drug administration by injection mode and possible safety issue, further improve quality of the pharmaceutical preparations level and safety and effectiveness, the inventor adopts modern preparation technique through studying repeatedly, develop oral Antihepatitis medicament by the reform of craft screening, optimization and dosage form, thereby finished the present invention.
The object of the invention provides a kind of more efficiently oral anti-hepatitis Chinese medicine extract and preparation method thereof.
Another object of the present invention provides a kind of anti-hepatitis Chinese medicinal tablet and preparation method thereof.
Medicine of the present invention is made by following materials of weight proportions medicine: Herba Artemisiae Scopariae 5～25kg, Fructus Gardeniae 2～10kg, Radix Scutellariae 0.5～5kg, Radix Et Rhizoma Rhei 1～5kg, Rhizoma Coptidis 1～5kg and Cortex Phellodendri 1～5kg.
In order to reach better therapeutic, the weight ratio of crude drug also can be Herba Artemisiae Scopariae 5.56kg, Fructus Gardeniae 3.70kg, Radix Scutellariae 1.85kg, Radix Et Rhizoma Rhei 1.85kg, Rhizoma Coptidis 1.85kg and Cortex Phellodendri 1.85kg.
Chinese medicine extract of the present invention is made up of the effective site of Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and the Radix Et Rhizoma Rhei of above-mentioned weight proportion and the effective site of Rhizoma Coptidis and Cortex Phellodendri.
Chinese medicinal tablet of the present invention comprise active component and or pharmaceutically acceptable additive of tablet, the effective site that effective site that wherein said active component is made by Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and the Radix Et Rhizoma Rhei of above-mentioned weight proportion and Rhizoma Coptidis and Cortex Phellodendri make is formed by 3.5: 1 weight ratio.Described additive of tablet can be selected carboxymethyl starch sodium for use, adds carboxymethyl starch sodium in active component and 3: 1 ratio of carboxymethyl starch sodium mass ratio, and the magnesium stearate of adding tablet total weight amount 0.3% is as lubricant.
Chinese medicine extract of the present invention and tablet have heat clearing away, detoxifcation, dampness removing, jaundice eliminating to hepatitis, the effect of acute due to being used for accumulateing in the damp and hot malicious heresy, delay property, chronic hepatitis, hepatitis gravis and liver cirrhosis jaundice and biliary tract infection, cholelithiasis etc.
The preparation method of Chinese medicine extract of the present invention is as follows:
Take by weighing crude drug Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae, Radix Et Rhizoma Rhei, Rhizoma Coptidis and the Cortex Phellodendri of above-mentioned weight ratio, this 6 flavor crude drug is divided into 2 groups, wherein Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei are one group, and Rhizoma Coptidis and Cortex Phellodendri are another group;
One, the yellow effective site (Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae, Radix Et Rhizoma Rhei) of mattress Cape jasmine a kind of reed mentioned in ancient books
(1) gets Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae, Radix Et Rhizoma Rhei, add 6～12 times in water, soak after 0.5～3 hour, Herba Artemisiae Scopariae and Fructus Gardeniae added in the entry decoct, boil the back and add Radix Scutellariae, add Radix Et Rhizoma Rhei again after boiling etc. decoction liquor is little, begin to calculate decocting time, 0.5 after～2 hours preferred 1 hour, decoction liquor is leached reservation, adding water in the vessel continues to decoct 2 times, each 0.5～2 hour preferred 1 hour, merge the decoction liquor that leaches, being evaporated to 80 ℃ of following relative densities is 1.05～1.10, and the mass volume ratio that obtains medical material and medicinal liquid is 1: 2～3 (W/V), concentration is the clear paste of 0.3～0.5 gram crude drug/milliliter, the clear paste placement is spent the night centrifugal filtration, collecting precipitation and filtrate respectively;
(2) filtrate with step (1) gained adds the good D101 macroporous adsorptive resins of pretreatment, choose blade diameter length ratio and be 1: 5～1: 9 preferred 1: 7; Medical material: resin is 1: 2～1: 4 preferred 1: 3, and the control flow velocity is 2～3 times of resin bed volumes per hour, treated that the post liquid stream to the greatest extent after, washing, being negative with the Molish reaction is washing judgement terminal point; Be the ethanol elution of 50%～90% ethanol preferred 70% then with concentration, therefore color is that pure eluting is judged terminal point to add the strong aqua ammonia permanent red in the eluent because Radix Et Rhizoma Rhei anthraquinone chance alkali reddens, collect pure washing liquid, the washing volume is 10 times of resin bed volume in this step, and 70% alcohol is washed volume and is about 10 times of resin bed volume;
(3) with the precipitation of gained in the step (1) with 6 times of amounts (W/V), 95% alcohol reflux 1 hour, filter, resinol washing liquid in filtrate and the step (2) merges, decompression recycling ethanol, being concentrated into 80 ℃ of following relative densities is 1.25～1.30, and 70 ℃ of vacuum dryings are pulverized then, cross 100 mesh sieves, promptly get the yellow effective site of mattress Cape jasmine a kind of reed mentioned in ancient books.
Two, Rhizoma Coptidis Cortex Phellodendri effective site
(1) getting Rhizoma Coptidis, Cortex Phellodendri, is alcohol reflux 1-5 time of 85%～95% ethanol preferred 90% with concentration, merges alcohol extract, and placement is spent the night, and draws supernatant, and reclaim under reduced pressure is not to there being the alcohol flavor, and placement is spent the night, and filtrate and precipitation are collected in centrifugal filtration respectively;
(2) water washing and precipitating, clear and bright to water lotion, precipitate standbyly, collect water lotion;
(3) filtrate of step (1) gained and the water lotion of step (2) gained are merged, transfer pH to 1～4 with concentrated hydrochloric acid, placement is spent the night, and separates out precipitation, and it is closely neutral that precipitation is washed to pH;
(4) precipitation with step (2) and step (3) gained merges, and 60-80 ℃ of following vacuum drying pulverized, and crosses the 50-200 mesh sieve, promptly gets Rhizoma Coptidis Cortex Phellodendri effective site (total alkaloids).
Merge Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae, Radix Et Rhizoma Rhei effective site and Rhizoma Coptidis, the Cortex Phellodendri effective site that above step obtains and promptly get Chinese medicine extract of the present invention.
Chinese medicine extract of the present invention can add the required various conventional adjuvant of preparation different dosage form, be prepared into any peroral dosage form commonly used as disintegrating agent, lubricant, binding agent etc. according to the method for Chinese medicinal and the supplementary product consumption of routine, as pill, powder, tablet, capsule etc.
The preparation process of mattress Cape jasmine a kind of reed mentioned in ancient books Huang effective site and Rhizoma Coptidis Cortex Phellodendri effective site is identical in the preparation process of yellow effective site of mattress Cape jasmine a kind of reed mentioned in ancient books in the Chinese medicinal tablet active component of the present invention and Rhizoma Coptidis Cortex Phellodendri effective site and the Chinese medicine extract, difference is: in the preparation process of tablet with the Herba Artemisiae Scopariae that makes, Fructus Gardeniae, Radix Scutellariae, Radix Et Rhizoma Rhei effective site and Rhizoma Coptidis, Cortex Phellodendri effective site evenly obtains active component of the present invention by 3.5: 1 mixed, with the active component of gained and carboxymethyl starch sodium in 3: 1 ratio of mass ratio, add carboxymethyl starch sodium as disintegrating agent and excipient, wherein 2/3 amount adds when granulating, 1/3 amount adds when tabletting, and concrete steps are as follows:
(1) granulates: take by weighing active component and carboxymethyl starch sodium in 3: 1 ratio of mass ratio according to active component and carboxymethyl starch sodium, and carboxymethyl starch sodium is divided into two fens, portion is that 1/3 amount is standby, another part is that 2/3 amount joins in the active component, with concentration is that 70%～80% ethanol is that wetting agent is granulated, and the granule that makes is dry down in 60 ℃;
(2) tabletting: get dried particles, add the carboxymethyl starch sodium of 1/3 standby amount and the magnesium stearate of tablet total weight amount 0.3%, depress to tablet with 1～10kN pressure.
Chinese medicine extract of the present invention and tablet have following effect:
1, choleretic effect can increase the choleresis of normal rat, and the inductive bile duct injury rat bile secretion of oral ANIT is increased, and shows that the clear pornographic movie of positive liver has tangible choleretic effect.
2, the jaundice eliminating effect can suppress the rising of serum bilirubin that oral ANIT causes the hepatic injury rat, cholesterol, cholic acid.Show its significant jaundice eliminating effect.
3, function for protecting liver and reducing enzyme activity can suppress multiple injection CCl
4The rat blood serum transaminase's who causes rising, total protein and albumin reduce, and reduce the liver collagen content; To disposable ip carbon tetrachloride (CCl
4), the rising of D-galactosamine (D-GalN) induced mice serum glutamic pyruvic transminase (GPT), glutamic oxaloacetic transaminase, GOT (GOT) also has stronger inhibitory action, histological examination is to respectively by CCl
4, the animal liver damage that causes of D-GalN, the positive clear pornographic movie of liver has the certain protection effect.
4, immune influence is added the inductive mouse immune liver damage of LPS (lipopolysaccharide) to BCG (bacillus calmette-guerin vaccine) significant protective effect is arranged; Can obviously strengthen ring phosphinylidyne ammonia (Cy) and handle low peritoneal macrophage phagocytic function, hemolytic antibody reaction of formation and the inductive lymphproliferation response of Con A of mice, and can strengthen the inductive lymphproliferation response of normal mouse Con A and show that the clear pornographic movie of positive liver has the effect of starving the enhancing human body immunity ability preferably.
5, the hepatitis virus resisting effect can suppress duplicating and expressing of 2.2.15 cell HBsAg, HBeAg, and dna replication dna has inhibitory action in pair cell culture supernatant and the cell; The clear middle DHBVDNA rising of Sanguis Anas domestica to infected duck hepatitis B virus (DHBV) has good inhibitory effect.The positive clear pornographic movie of liver has certain inhibitory action to hepatitis B virus.
The specific embodiment:
Below further set forth beneficial effect of the present invention by pharmacodynamics test.
Experiment material:
1, be subjected to the reagent thing: Chinese medicine extract tablet of the present invention (calling " the positive clear pornographic movie of liver " in the following text), the 302nd hospital of PLA provides, lot number 010620.
2, positive control drug: Ursofalk, German good fortune restrains big pharmaceutical factory, the 99J15043L01 bifendate, the Beijing XieHe medicine Factory, lot number: 010309, levamisole hydrochloride, Hunan Dongting Pharmaceutical Co., Ltd. produces, lot number 010832.Lamivudine, Britain Glaxo Wellcome company limited B022306.
3, reagent: glutamate pyruvate transaminase (GPT), glutamic oxaloacetic transaminase, GOT (GOT) are measured test kit, are the safe clinical reagent company limited of Beijing northization product.Total bilirubin (TBil), cholesterol (CHE), total protein (TP), albumin (ALB) are measured test kit, are Beijing Zhongsheng Biological Engineering High Technology Company's product.The bile acid determination reagent box is a German Human company product.Other reagent is commercially available analytical pure.Cyclophosphamide (Cy) injectable powder, Hengrui Medicine Co., Ltd., Jiangsu Prov. produces, lot number 01060821.D-galactosamine (D-GalN), α-ANIT (ANIT) analytical pure, chemical defence institute of the Chinese People's Liberation Army provides.Bacillus calmette-guerin vaccine (BCG), Beijing Biological Product Inst. produces, lot number 010910.Lipopolysaccharide (LPS), Sigma company product, lot number L2688.Phytohaemagglutinin (PHA), Sigma company product, lot number L8754.Eagles MEM dry powder, G-418 (Geneticin), yeast t-RNA, E.C. 3.4.21.64, U.S. GIBCO company product; Hyclone, U.S. Hyclone Lab company product; L-glutaminate, the import packing of Jing Ke chemical reagents corporation; HBsAg, HBeAg solid phase ria-determination box, Beifang Inst. of Immune Reagents, Chinese Isotopes Co.; Kanamycin, North China Pharmaceutical Factory's product; Polyethylene Glycol, Sweden Fluka product; D-
32P-dCTP, the inferior brightness biomedical engineering in Beijing company product.
4, animal: the Wistar rat, body weight 190-210g, Kunming mouse, body weight 18-22g, all male and female half and half are provided by Military Medical Science Institute's animal center, and the animal quality certification number is respectively the moving word BDW98012 of medical officer and the moving word of medical officer BDW95001 number.1 age in days Beijing duck, 80-100g, planting institute animal feeding field by Beijing medical courses in general institute medicine provides.The zoopery condition: two grade standards, the quality certification number: medical officer moves word B98006.
5, DHB (DHBV-DNA) strong positive serum picks up from Shanghai and infects hepatitis B virus sheldrake ,-70 ℃ of preservations.
6, the 2.2.15 cell is the human liver cancer cell (HepG2) of clone's transfection hepatitis B viruses (HBV) DNA, and U.S. Mount Sinai medical center makes up, the cultivation of going down to posterity of medical biotechnology institute of Chinese Academy of Medical Sciences Viral Laboratory.
7, instrument: BS-224 type biochemical analysis analyzer, Beijing biochemical instrument factory.Hervester96 type 96 orifice plate cell harvesting instrument, U.S. TOMTE company produces.Instrument in the MicroBetu Trilux1450 type trace liquid, Perkin Elmer company produces.
Experimental technique and result:
One, the experimentation of the positive clear pornographic movie choleretic effect of liver
1, the positive clear pornographic movie of liver is tested the function of gallbladder promoting of normal rat: get the Wistar rat, be divided into 5 groups at random by body weight, every group 10, normal control group, Ursofalk group 0.135g/kg day, positive three dosage groups of the clear pornographic movie of liver high, medium and low (0.576g, 0.288g, 0.144g/kg day).Self administration of medication beginning in the 6th day, respectively 24,48,72,96,120 hours five time periods, (ketamine: anesthetized rat diazepam=1: 1), the separation ductus choledochus inserts the bile collecting pipe, collects 4 hours bile to adopt combined anesthesia.Calculate each time period, every animal, every 100g body weight bile flow.
The positive clear pornographic movie of liver of table 1 is to the excretory influence of normal rat bile (n=10)
Compare with the normal control group,
*P＜0.01,
*P＜0.05
2, the positive clear pornographic movie of liver is got the Wistar rat to the function of gallbladder promoting experiment of ANIT hepatic injury rat, be divided into 6 groups at random by body weight, every group 10, normal control group, liver injury model group, Ursofalk group 0.135g/kg day, positive three dosage groups of the clear pornographic movie of liver high, medium and low (0.576g, 0.288g, 0.144g/kg day).Beginning in the 6th day behind the self administration of medication, except that the normal control group, other each group is irritated stomach with 4%ANIT oil preparation 1.75ml/kg, cause rat bile alluvial model, respectively 24,48,72,96,120 hours five time periods behind the modeling type, adopt combined anesthesia (ketamine: anesthetized rat diazepam=1: 1), separation ductus choledochus, insert the bile collecting pipe, collect 4 hours bile.Calculate each time period, every animal, every 100g body weight bile flow.
The positive clear pornographic movie of liver of table 2 is to the excretory influence of ANIT hepatic injury rat bile (n=10)
Compare #P＜0.05 with the normal control group
##Compare according to group with model P＜0.01
*P＜0.05
*P＜0.01
Table 1,2 results show that the positive clear pornographic movie of liver can make normal rat, the secretion of ANIT hepatic injury rat bile increase, and points out the clear pornographic movie of positive liver to have choleretic effect.
Two, the jaundice eliminating of the positive clear pornographic movie of liver experiment: get the Wistar rat, be divided into 6 groups at random by body weight, every group 10, normal control group, liver injury model group, Ursofalk group 0.135g/kg day, positive three dosage groups of the clear pornographic movie of liver high, medium and low (0.576g, 0.288g, 0.144g/kg day).Beginning in the 6th day behind the self administration of medication, except that the normal control group, other each group is irritated stomach with 4%ANIT oil preparation 1.75ml/kg, causes rat bile alluvial model, respectively 24,48,72,96,120 hours five time periods after moulding, gets rat femoral blood.Measure total bilirubin (TBil), cholesterol (CHE), cholic acid value, and get hepatic tissue and do the pathology inspection.
The positive clear pornographic movie agent of liver of table 3 is to the influence (n=10) of rat blood serum mesobilirubin
Compare with the normal control group
#P＜0.05
##Compare with model group P＜0.01
*P＜0.05
*P＜0.01
The positive clear pornographic movie agent of liver of table 4 is to the influence (n=10) of cholesterol in the rat blood serum
Compare with the normal control group
#P＜0.05
##Compare with model group P＜0.01
*P＜0.05
*P＜0.01
The positive clear pornographic movie agent of liver of table 5 is to the influence (n=10) of cholic acid in the rat blood serum
Compare with the normal control group
*P＜0.05
** P＜0.05 * * P＜0.01 is compared with model group in P＜0.01
Three. the liver protecting and ALT lowering experiment of the positive clear pornographic movie of liver
1, carbon tetrachloride is caused the influence of rat chronic hepatic injury: get the Wistar rat, be divided into 6 groups at random by body weight, every group 20, normal control group, liver injury model group, bifendate group, the positive clear pornographic movie of liver high, medium and low (0.576g, 0.288g, 0.144g/kg the day/kg) three dosage groups.Except that the normal control group, each treated animal lumbar injection 10%CCl
4Vegetable oil 0.5ml/100g body weight, weekly twice, totally 12 weeks, normal control group ip normal saline.Every day, ig was administered once (1ml/100g body weight, normal control group ig is with the distilled water of volume) simultaneously in modeling.Before experiment finished, animal fasting 12 hours was plucked the eyeball blood sampling with animal, with 3000 rev/mins centrifugal 10 minutes, separation of serum is measured GPT, GOT, TP, ALB, with sacrifice of animal, get hepatic tissue and make hydroxyproline determination after the blood sampling, calculate liver collagen content (the results are shown in Table 6,7).
The positive clear pornographic movie of liver of table 6 causes the influence of rat chronic hepatic injury to carbon tetrachloride
Compare with normal group
*P＜0.05
*Compare with model group p＜0.01
*P＜0.05
*P＜0.01
The positive clear pornographic movie of liver of table 7. is to the influence of rats'liver collagen content (X ± SD)
Compare * p＜0.05 * * p＜0.01 with model group
Table 6,7 results show that the positive clear pornographic movie of liver can suppress multiple injection CCl
4The rat blood serum transaminase's who causes rising, total protein and albumin reduce, and reduce the liver collagen content.
2, to the influence of D-GalN induced mice acute liver damage: get 60 of healthy mices and be divided into six groups at random, be three dosage groups of normal control group, D-GalN (800mg/kg) model group, bifendate group 0.029g/kg, the positive clear pornographic movie of liver (0.832,0.416,0.208g/kg), 10 every group.Every day, the ig administration was 2 times, and matched group and model group are given the distilled water with same volume, after the administration 7 times, and except that the normal control group, ip in mice D-GalN 0.2ml/10g, after the modeling 20 hours, mice socket of the eye venous blood collection was surveyed serum GPT, GOT value (table 8).
The positive clear pornographic movie of liver of table 8. causes the influence (X ± SD) of chmice acute hepatic injury to D-GalN
Compare with the normal control group,
#P＜0.05
##Compare with model group p＜0.01,
*P＜0.05
*P＜0.01
Table 8 is the result show, each administration group mice serum GPT, GOT value all are lower than model control group, the positive clear pornographic movie height of liver, middle dosage group and model group relatively have significant difference, illustrate that the clear pornographic movie of positive liver has certain therapeutical effect to D-GalN induced mice acute liver damage.
3, to CCl
4The influence of induced mice acute liver damage: get 60 of healthy mices and be divided into six groups at random, be i.e. normal control group, CCl
4(0.02ml/kg) three dosage groups of model group, bifendate group 0.029g/kg, the positive clear pornographic movie of liver (0.832,0.416,0.208g/kg), 10 every group.Every day, the ig administration was 2 times, and matched group and model group are given the distilled water with same volume, behind 7 medicines of successive administration, and except that the normal control group, ip in mice 0.1%CCl
40.2ml/10g after the modeling 40 hours, mice socket of the eye venous blood collection was surveyed serum GPT, GOT value (seeing Table 9).
The positive clear pornographic movie of liver of table 9. causes the influence (X ± SD) of chmice acute hepatic injury to CC14
Compare with the normal control group, * p＜0.05 * * p＜0.01 is compared with model group in #p＜0.05 ##p＜0.01
Table 9 is the result show, each administration group rat blood serum GPT, GOT value all are lower than model control group, and the positive clear pornographic movie height of liver, middle dosage group and model group relatively have significant difference, illustrate that the clear pornographic movie of positive liver is to CCl
4The induced mice acute liver damage has therapeutical effect preferably.
Four, the positive clear pornographic movie sheet of liver is to effect of immunologic function
1, to the influence of immunologic liver injury: 60 of healthy mices, be divided into six groups at random, normal control group, model group, bifendate group (0.029g/kg), positive three dosage groups of the clear pornographic movie of liver (0.832g/kg, 0.416g, 0.208g), 10 every group, mice ig administration 2 times/day, totally 10 days.Administration is preceding except that the normal control group, every caudal vein injection of BCG 5 * 10
7U/, every Mus tail vein injection LPS 7.5 μ g attack again after 10 days, the eyeball rear vein beard is got blood and is surveyed serum GPT, GOT after 12 hours.The results are shown in Table 10.
The positive clear pornographic movie sheet of liver of table 10. is to the influence of mouse immune liver damage (X ± SD)
Compare with matched group
##* p＜0.05 * * p＜0.01 is compared with model group in p＜0.01
The result shows, the positive clear pornographic movie of liver has the obvious suppression effect to the rising of immunologic liver injury mice serum transaminase due to the BCG+LPS, with model group significant difference is arranged relatively.
2, to the influence of Turnover of Mouse Peritoneal Macrophages phagocytic function: chicken red blood cell is engulfed method.Get healthy male mice, be divided into six groups at random, three dosage groups of the left-handed rice of normal control group, cyclophosphamide (Cy) model group, Cy+ positive control drug azoles group (0.04g/kg), the clear pornographic movie of the positive liver of Cy+ (0.832,0.416,0.208g/kg), every group 10, gastric infusion (ig), each 0.2ml/, every day 1 time, in a continuous week, normal control group and Cy model group give the equivalent distilled water.Be subjected to reagent thing 1h pneumoretroperitoneum injection (ip) Cy15mg/kg in the 4th day and the 5th day ig, 1 time/day, the normal control group is given distilled water with method.All animals administers are in the time of the 6th day, lumbar injection 5% starch normal saline solution, and the 1mL/ Mus is to stimulate the gathering and the activation of peritoneal macrophage.Tested preceding 45 minutes, lumbar injection 5% chicken erythrocyte suspension 0.4mL/ Mus, disconnected ridge is put to death mice, the abdominal cavity injects normal saline 1mL/ Mus, gently rub abdominal part, open the abdominal cavity, get the about 0.2mL of peritoneal fluid and drip on clean slide, put 37 ℃ and incubate temperature and take out slide after 30 minutes, use the normal saline rinsing, remove not paster cell, dry the back and fix 5 minutes with 1: 1 acetone-methanol solution, Ji Mu Sa-Rui Shi mixed liquor dyeed 5 minutes, and washing is dried, the oil mirror is engulfed the macrophage number of chicken red blood cell and is engulfed the sum of chicken red blood cell in per 100 macrophages of counting down, and is calculated as follows phagocytic percentage and phagocytic index.
The positive clear pornographic movie of liver of table 11 is handled the influence (X ± SD) of Turnover of Mouse Peritoneal Macrophages phagocytic function to Cy
Compare with the normal control group
##Compare with the Cy model group p＜0.01
*P＜0.01,
By table 11 result as seen, phagocytic percentage and the normal matched group of phagocytic index that Cy model mice peritoneal macrophage is engulfed chicken red blood cell obviously reduce, give high, medium and low three dosage of the clear pornographic movie of positive liver and positive control drug assistant and revolve imidazoles phagocytic percentage and phagocytic index are obviously raise, point out the clear pornographic movie of positive liver that the reduction tool of Cy induced mice peritoneal macrophage phagocytic function is improved significantly.
3, the influence that the mice hemolytic antibody is generated: get healthy male mice, be divided into six groups at random by body weight, three dosage groups of normal control group, cyclophosphamide (Cy) model group, Cy+ positive control drug levamisole group, the clear pornographic movie of the positive liver of Cy+ (0.832,0.416,0.208g/kg), 10 every group.Ig administration every day 1 time (the isopyknic distilled water of normal control group ig), continuous 7 days, behind the medicine the 5th day, sheep red blood cell (SRBC) (SRBC) the suspension 0.2ml/ of every Mus ip (V/V) dilution in 3: 5 only carries out immunity, after one hour, except that the normal control group, each organizes ip in mice cyclophosphamide 15mg/kg body weight (0.2ml/20g), and next day is again to inject once with dosage.After the immunity four days, extract eyeball of mouse and get blood, separation of serum, with normal saline serum is diluted 200 times, get the 1ml dilute serum and add 10%0.5ml SRBC, put in the ice bath, every pipe adds the guinea pig serum that 1ml diluted with normal saline at 1: 10, moves to immediately in 37 ℃ of waters bath with thermostatic control, be incubated 10 minutes, promptly put into ice bath with cessation reaction,, get supernatant 1ml with the centrifugal 10min of 2000r/min, add 3ml Dou Shi liquid mixing, placing after 10 minutes, is blank with the control tube, measures trap (OD) value of each sample under 540nm with spectrophotometer.Other gets SRBC diluent 0.125ml, adds Dou Shi liquid 2ml, puts and measures OD value (540nm), trap value when this OD value (540nm) is the SRBC HD50 after 10 minutes.Calculate the half hemolysis value (HC of every sample by following formula
50).
The serum hemolysin reaction system
The positive clear pornographic movie of liver of table 12 is handled the influence that the mice hemolytic antibody generates to Cy
Compare with the normal control group ##P＜0.01; Compare with the Cy model group * P＜0.01
The results are shown in Table 12, the positive clear pornographic movie of liver can obviously increase Cy model mice serum hemolysin content, and immunoreation due to the Cy is suppressed to have stronger antagonism.
4, to the influence of mouse lymphocyte breeder reaction: get the Healthy female mice, be divided into six groups at random by body weight, three dosage groups of normal control group, Cy model group, Cy+ positive control drug levamisole group, the clear pornographic movie of the positive liver of Cy+ (0.832,0.416,0.208g/kg), 10 every group.Ig administration every day 1 time (normal control group and the isopyknic distilled water of Cy model group ig), continuous 7 days.Behind the medicine the 5th day, except that the normal control group, each organized ip in mice cyclophosphamide 15mg/kg body weight (0.2ml/20g), and next day is again with the dosage injection once.Got mouse spleen on the 8th day and prepare splenocyte suspension, dye exclusion method identification of cell survival rate is more than 95%.Use methyl
3H～thymidine (
3H-TdR) method of mixing is measured the influence of the clear pornographic movie of positive liver to the inductive lymphproliferation response of mice Con A.
3H-TdR mixes method: splenocyte concentration is adjusted into 5 * 10
6/ ml, add splenocyte suspension 100 μ l/ holes to Costar 96 orifice plates, the Con A 100 μ l/ holes (detecting the spontaneous breeder reaction of the inductive splenocyte of Con A) that add 1640 liquid, 100 μ l/ holes (detecting the spontaneous breeder reaction of splenocyte) or 1 μ g/ml then, if 37 ℃, saturated humidity and 5%CO are put in 10 multiple holes
2Cultivate 7 2h in the incubator, 16h adds 10 μ Ci/ml's before stopping cultivation
3H-TdR20 μ l (7.4KBq)/hole.Collect sample on filter membrane with the Hervester 96 types 96 orifice plate cell harvesting instrument that U.S. TOMTE company produces, add scintillation solution after the drying and measure every hole radioactivity (cpm) with instrument in the MicroBetu Trilux of the Perkin Elmer company 1450 types trace liquid.The result is with the average cpm value representation in average 10 multiple holes.
The positive clear pornographic movie of liver of table 13 is handled the influence of mouse boosting cell breeder reaction to Cy
Compare with the normal control group ##P＜0.01; Compare with the Cy model group * P＜0.01
5, external influence to the mouse lymphocyte breeder reaction: get 3 healthy adult female mice spleens, conventional method prepares single cell suspension, with 1640 liquid diluting cells to 5 * 10 of 20% calf serum
6/ ml adds cell suspension 100 μ l in the every hole of 96 porocyte culture plates, Con A (1.0ug/ml) 50 μ l, and the medicine 50 μ l of variable concentrations, control wells adds isopyknic 1640 liquid, and every sample is all established 6 multiple holes, puts 37 ℃, saturated humidity and 5%CO
2Incubate and cultivate 56h in the incubator, every then hole adds 10 μ Ci/ml's
3H-TdR 20 μ l continue to cultivate 16h.Collect sample on filter membrane with the Hervester 96 types 96 orifice plate cell harvesting instrument that U.S. TOMTE company produces, add scintillation solution after the drying and measure every hole radioactivity (cpm) with instrument in the Perkin Elmer MicroBetuTrilux of the company 1450 types trace liquid.The result is with the average cpm value representation in average 6 multiple holes.
The positive external influence of the clear pornographic movie of liver (n=6) of table 14 to the mouse boosting cell breeder reaction
Compare #P＜0.05 with matched group, ##P＜0.01,
The result is as shown in table 14, and just the clear pornographic movie of liver is external can increase splenocyte in the 0.5-50.0ug/ml dosage range
3The H-TdR value of mixing, pointing out the clear pornographic movie of positive liver, external breeder reaction has potentiation to mouse lymphocyte.
6, to the influence of mice circulating immune complex (CIC) level: Polyethylene Glycol (PEG) direct precipitation method.Get the Healthy female mice, be divided into six groups at random, three dosage groups of normal control group, ring phosphinylidyne ammonia (Cy) model group, Cy+ positive control drug levamisole group, the clear pornographic movie of the positive liver of Cy+ (0.832,0.416,0.208g/kg), 10 every group by body weight.Ig administration every day 1 time (normal control group and the isopyknic distilled water of Cy model group ig), continuous 7 days.Behind the medicine the 5th day, except that the normal control group, each organized ip in mice cyclophosphamide 15mg/kg body weight (0.2ml/20g), and next day is again with the dosage injection once.Extract eyeball of mouse on the 8th day and get blood, separation of serum, application of sample is in 96 orifice plates, every sample is established control wells and experimental port, control wells is the borate 200 μ l+ mice serums 22 μ l of 0.1mol/L pH 8.4, experimental port is 7.5%PEG 200 μ l+ mice serums 22 μ l, and room temperature is placed 1h, measures OD value (450nm) with porous scanning spectrophotometer Multiskan Mce/340 MK II.The result is with experimental port OD value-control wells OD value representation.
The positive clear pornographic movie of liver of table 15 is handled the influence of mice circulating immune complex level to Cy
Compare with the normal control group ##P＜0.01; * P＜0.05, * * P＜0.01, with the Cy model group relatively
The result is as shown in Table 15, the normal matched group of Cy model mice circulating immune complex level raises, give high and low two dosage of the clear pornographic movie of positive liver and positive control drug assistant and revolve imidazoles the circulating immune complex level is reduced, point out the clear pornographic movie of positive liver that Cy induced mice circulating immune complex level rising tool is had some improvement.
The positive external influence of the clear pornographic movie of liver of table 16 to the mouse boosting cell breeder reaction
Compare with matched group
*P＜0.05,
*P＜0.01,
The result: the positive clear pornographic movie of liver can obviously strengthen Cy and handle low peritoneal macrophage phagocytic function, hemolytic antibody reaction of formation and the inductive lymphproliferation response of Con A of mice in the 0.028-0.832g/ml dosage range, shows that the clear pornographic movie of positive liver has obvious improvement effect to immunologic hypofunction due to the Cy.In addition, the positive external inductive lymphproliferation response of mice Con A that in the 0.5-50.0ug/ml dosage range, also can strengthen of the clear pornographic movie of liver.
Five, the clear pornographic movie anti-hepatitis virus experiment of positive liver
1, in the duck body to the influence of DHB: 1 age in days Beijing duck, clear through the positive Sanguis Anas domestica of lower limb shin intravenous injection Shanghai sheldrake DHBV-DNA, every 0.3ml infects and got blood in back 7 days, separation of serum detects DHBVDNA content in the serum.Duckling serum is divided into 5 groups at random with duck after DHBV is positive after testing, three dosage groups of virus control group, lamivudine group (50mg/kg), the positive clear pornographic movie of liver (8,4,2g/kg), 6 every group.Gastric infusion, 1.5ml/, every day 2 times.Matched group gives with the volume normal saline, successive administration 10 days.Respectively with Drug therapy after 5 days (T
5), 10 days (T
10) and drug withdrawal after 3 days (P
3) get blood from duck shin vein, separation of serum is pressed nick translation test kit description method, uses
32P labelling DHBVDNA probe, and make the clear dot blot hybridization of Sanguis Anas domestica, autoradiography diaphragm speckle, microplate reader is measured OD value (optical filter is 490nm), calculating serum DHBV-DNA optical density, with hybridization spot OD value as specimen DHBV-DNA level value.The results are shown in Table 17, table 18.
The positive clear pornographic movie of liver of table 17. in the duck body to the influence of DHB DHBV-DNA
Statistical disposition: different time (T after the administration
5, T
10, P
3) (T before OD value and the administration on the same group
0) OD value comparison (paired t-test).
*p＜0.05，**p＜0.01，***p1＜0.001。
Positive clear yellow treatment group of liver of table 18. and the horizontal rejection ratio of the clear DHBV-DNA of viral infection matched group Sanguis Anas domestica are
Statistical disposition: drug treatment group HDV-DNA suppression ratio and virus control group compare (t check in groups) with the DHBV-DNA suppression ratio of time.
*P＜0.05，
**P＜0.01，
***P＜0.001。
Three batches of experimental results show that one day 2 times 10 days effects always to the hepatitis b virus infected clear middle DHBVDNA level of Sanguis Anas domestica of infected duck of the positive clear yellow 8.0g/kg of liver have significance effect, non-toxic reaction; 4.0g/kg group administration the 5th day in second batch and the 3rd batch of test had certain inhibitory action on the tenth day.The oral inhibitory action to DHBVDNA of positive drug lamivudine is extremely remarkable.Above experimental result shows: the clear Huang of positive liver has therapeutical effect preferably to the duck hepatitis B virus infection duck.
2, in the 2.2.15 cell to hepatitis B virus surface antigen, the antigenic inhibitory action of e: the clear pornographic movie of the positive liver of medicine is mixed with culture medium and contains crude drug 10mg/L solution, with 2 times be diluted to 5,2.5,1.25,0.625,0.3125,0.156.3mg/ml adds 96 porocyte culture plates, every concentration 4 holes, change same concentration liquid in per 4 days, and established no medicine cell matched group.With the observation of cell pathological changes is index, 8 days microscopically observation of cell pathological changes, and destroying fully is 4; 75% is 3; 50% is 2; 25% is 1; Anosisly become 0.Calculate every concentration liquid average cell lesion degree and suppress percentage rate.Press Reed Meuench method and calculate TC
50, TC
0, see Table 19.
(A=log＞50% suppresses percentage rate B=log＜50% drug level C=log extension rate)
The toxicity of the clear Huang of the positive liver of table 19 pair cell in the 2.2.15 cell culture
Table 19 shows, it is 1.2mg/ml that the positive clear pornographic movie of liver produces 50% inhibitory action concentration to the 2.2.15 cell, avirulence concentration is 0.6mg/ml, points out the clear pornographic movie of positive liver should select 0.6mg/ml to the drug effect concentration of 2.2.15 cell, and at these 2 times of dilution test medicinal liquids below concentration.
With 100,000 every milliliter 2.2.15 cell inoculation 24 well culture plates, every hole 1ml, 37 ℃ of 5%CO
2Cultivated 24 hours, and added the following 2 times of dilution test medicinal liquids of non-toxic concn, 5 dilution factors are respectively 4,2,1,0.5,0.25mg/ml, every concentration 3 holes, 37 ℃ of 5%CO
2Cultivate, collect the 8th day pastille culture fluid ,-20 ℃ of stored frozen.Measure with reference to solid phase radioimmunoassay box description, measure every hole cpm value with the γ calculating instrument.HBsAg, the HBeAg positive and negative control are established in experiment.See Table 20,21.
The positive clear pornographic movie of liver of table 20 in the 2.2.15 cell to the influence (two batches of experimental results) of HBsAg
Compare * * p＜0.01 * p＜0.05 with cell
The positive clear pornographic movie of liver of table 21 in the 2.2.15 cell to the influence (three batches of experimental results) of HBeAg
Compare * * p＜0.01 * p＜0.05 with cell
3, positive clear Huang of liver and lamivudine are to the inhibitory action of 2.2.15 cell culture supernatant HBV-DNA
3.1, positive liver clear yellow in the 2.2.15 cell culture supernatant HBV-DNA dot blot hybridization: get 1,000,000 every milliliter inoculations of 2.2.15 cell 24 porocyte culture plates, every hole 1ml, inoculate back 24 hours and add medicine, changing the original content medicinal liquid in per 4 days cultivates, cultivate after the dosing and collected supernatant on the 8th day, through polyethylene glycol precipitation, E.C. 3.4.21.64 cracking, phenol: chloroform: steps such as isoamyl alcohol extracting, dehydrated alcohol precipitate nucleic acids, vacuum is drained, and heavily is dissolved in the TE buffer making sample.Get 20ul (dna content 25ug), through degeneration, neutralization, and with 20X SSC buffer two-fold dilution to 1: 8 times are diluted on the nitrocellulose filter, and, prehybridization roasting through doing, hybridize, wash steps such as film, autoradiography.With conventional method towards X-ray sheet developing.The scanner scanning mating plate is measured density with gel-pro software, calculates suppression ratio and IC50.See Table 22.
The positive liver of table 22. clear yellow in the 2.2.15 cell culture supernatant to the effect of HBV-DNA
First experiment HBV-DNA spot density value/suppression ratio %:
Second crowd of experiment HBV-DNA speckle relative density value/suppression ratio %
3.2, positive liver clear yellow in the 2.2.15 cell to the Southern Blot inhibitory action of HBV-DNA: cultivated 8 days after getting the dosing of 2.2.15 cell, absorb culture fluid and collect cell, cell is through the lysate cracking, equal-volume phenol: chloroform: isoamyl alcohol extracting 2 times, add the dehydrated alcohol precipitate nucleic acids, vacuum is drained, and heavily is dissolved in the 20ul TE buffer, add the DNA sample buffer, sample is added on electrophoresis on the agarose gel.Behind the electrophoresis successively behind degeneration, neutralization, commentaries on classics film.Together bake film, hybridization, exposure with the dot blot hybridization film.The scanner scanning mating plate with gel-pro gel analysis software analysis relative density, calculates suppression ratio and IC50.Experimental result sees Table 23.
The inhibition data of the clear Huang of the positive liver of table 23. HBV-DNA Southern Blot in the 2.2.15 cell
Experiment shows: the clear yellow median toxic concentration (TC50) of positive liver is 1200 μ g/ml.Maximal non-toxic concentration liquid (600 μ g/ml).Added the 2.2.15 cell culture 8 days, and obviously suppressed the secretion of HBsAg and HBeAg, IC50 is 400.95 ± 26.1 μ g/ml and 542.6 ± 76.5, and selection index is 2.99 and 2.21.The IC50 of HBV-DNA Southern Blot in the pair cell is 215.96 ± 8.88 μ g/ml, and selection index is 5.56.The IC50 of pair cell culture supernatant and intracellular HBV-DNA dot blot hybridization and Southern Blot is respectively 142.24 ± 13.04 and 156.98 ± 80.50 μ g/ml, and selection index is respectively 8.42 and 7.64.Results suggest: positive liver is clear yellow 2,2, in 15 cell culture hepatitis B virus is had inhibitory action.
Below by specific embodiment the preparation process of medicine of the present invention is described:
Embodiment 1: the preparation of Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Et Rhizoma Rhei and Radix Scutellariae effective site and Rhizoma Coptidis and Cortex Phellodendri effective site among the present invention:
Embodiment 1-1:
Get Herba Artemisiae Scopariae 5.56kg, Fructus Gardeniae 3.70kg, Radix Scutellariae 1.85kg, Radix Et Rhizoma Rhei 1.85kg, Rhizoma Coptidis 1.85kg and Cortex Phellodendri 1.85kg, standby;
One, Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Et Rhizoma Rhei and the preparation of Radix Scutellariae effective site
Get Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Et Rhizoma Rhei and Radix Scutellariae and add 6 times of water loggings bubble after 1 hour, Herba Artemisiae Scopariae and Fructus Gardeniae are added in the entry decoct, boil the back and add Radix Scutellariae, add Radix Et Rhizoma Rhei again after boiling etc. decoction liquor is little, begin to calculate decocting time, after 1 hour, decoction liquor is leached reservation, in vessel, add water continuation decoction 2 times, each 1 hour, merge the decoction liquor that leaches, being evaporated to relative density is 1.05, placement is spent the night, centrifugal filtration, collecting precipitation and filtrate respectively;
Filtrate is added the good D101 macroporous adsorptive resins of pretreatment, and selecting blade diameter length ratio is 1: 5, and medical material is 1: 2 with the ratio of resin, and flow velocity is 2 times of resin column volumes per hour, the upper prop liquid stream to the greatest extent after, washing, being negative with the Molish reaction is the judgement terminal point; Again with 50% ethanol elution, add to the eluent till the strong aqua ammonia permanent red, collect pure washing liquid;
To precipitate with 95% alcohol reflux of 6 times of amounts (W/V) 1 hour, filter, filtrate and resinol washing liquid will be merged, decompression recycling ethanol, being concentrated into 80 ℃ of following relative densities is 1.25,70 ℃ of vacuum dryings, pulverizes, mistake 100 mesh sieves obtain effective site, and are standby;
Two, Rhizoma Coptidis and Cortex Phellodendri effective site preparation:
Get Rhizoma Coptidis and Cortex Phellodendri, with 85% alcohol reflux 2 times, merge alcohol extract, placement is spent the night, and centrifugal filtration keeps filtrate, and water washing and precipitating is clear and bright to water lotion, precipitates standby;
Filtrate and water lotion are merged, regulate pH to pH=1 with concentrated hydrochloric acid, standing over night is separated out precipitation, and water washing and precipitating is to neutral;
Merge the gained precipitation 2 times, 60 ℃ of vacuum dryings are pulverized, and cross 70 mesh sieves, and obtaining total alkaloids is Rhizoma Coptidis and Cortex Phellodendri effective site.
Embodiment 1-2:
Get Herba Artemisiae Scopariae 15kg, Fructus Gardeniae 10kg, Radix Scutellariae 5kg, Radix Et Rhizoma Rhei 5kg, Rhizoma Coptidis 10kg and Cortex Phellodendri 10kg, standby;
One, Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Et Rhizoma Rhei and the preparation of Radix Scutellariae effective site
Get Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Et Rhizoma Rhei and Radix Scutellariae and add 9 times of water loggings bubble after 2 hours, Herba Artemisiae Scopariae and Fructus Gardeniae are added in the entry decoct, boil the back and add Radix Scutellariae, add Radix Et Rhizoma Rhei again after boiling etc. decoction liquor is little, begin to calculate decocting time, after 2 hours, decoction liquor is leached reservation, in vessel, add water continuation decoction 2 times, each 2 hours, merge the decoction liquor that leaches, being evaporated to relative density is 1.08, placement is spent the night, centrifugal filtration, collecting precipitation and filtrate respectively;
Filtrate is added the good D101 macroporous adsorptive resins of pretreatment, and selecting blade diameter length ratio is 1: 7, and medical material is 1: 3 with the ratio of resin, and flow velocity is 3 times of resin column volumes per hour, the upper prop liquid stream to the greatest extent after, washing, being negative with the Molish reaction is the judgement terminal point; Again with 70% ethanol elution, add to the eluent till the strong aqua ammonia permanent red, collect pure washing liquid;
To precipitate with 95% alcohol reflux of 6 times of amounts (W/V) 1 hour, filter, filtrate and resinol washing liquid will be merged, decompression recycling ethanol, being concentrated into 80 ℃ of following relative densities is 1.28,70 ℃ of vacuum dryings, pulverizes, mistake 100 mesh sieves obtain effective site, and are standby;
Two, Rhizoma Coptidis and Cortex Phellodendri effective site preparation:
Get Rhizoma Coptidis and Cortex Phellodendri, with 90% alcohol reflux 3 times, merge alcohol extract, placement is spent the night, and centrifugal filtration keeps filtrate, and water washing and precipitating is clear and bright to water lotion, precipitates standby;
Filtrate and water lotion are merged, regulate pH to pH=3 with concentrated hydrochloric acid, standing over night is separated out precipitation, and water washing and precipitating is to neutral;
Merge the gained precipitation 2 times, 70 ℃ of vacuum dryings are pulverized, and cross 100 mesh sieves, and obtaining total alkaloids is Rhizoma Coptidis and Cortex Phellodendri effective site.
Embodiment 1-3:
Get Herba Artemisiae Scopariae 25kg, Fructus Gardeniae 10kg, Radix Scutellariae 5kg, Radix Et Rhizoma Rhei 5kg, Rhizoma Coptidis 5kg and Cortex Phellodendri 5kg, standby;
One, Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Et Rhizoma Rhei and the preparation of Radix Scutellariae effective site
Get Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Et Rhizoma Rhei and Radix Scutellariae and add 12 times of water loggings bubble after 3 hours, Herba Artemisiae Scopariae and Fructus Gardeniae are added in the entry decoct, boil the back and add Radix Scutellariae, add Radix Et Rhizoma Rhei again after boiling etc. decoction liquor is little, begin to calculate decocting time, after 2 hours, decoction liquor is leached reservation, in vessel, add water continuation decoction 2 times, each 2 hours, merge the decoction liquor that leaches, being evaporated to relative density is 1.10, placement is spent the night, centrifugal filtration, collecting precipitation and filtrate respectively;
Filtrate is added the good D101 macroporous adsorptive resins of pretreatment, and selecting blade diameter length ratio is 1: 9, and medical material is 1: 4 with the ratio of resin, and flow velocity is 3 times of resin column volumes per hour, the upper prop liquid stream to the greatest extent after, washing, being negative with the Molish reaction is the judgement terminal point; Again with 90% ethanol elution, add to the eluent till the strong aqua ammonia permanent red, collect pure washing liquid;
To precipitate with 95% alcohol reflux of 6 times of amounts (W/V) 1 hour, filter, filtrate and resinol washing liquid will be merged, decompression recycling ethanol, being concentrated into 80 ℃ of following relative densities is 1.30,70 ℃ of vacuum dryings, pulverizes, mistake 100 mesh sieves obtain effective site, and are standby;
Two, Rhizoma Coptidis and Cortex Phellodendri effective site preparation:
Get Rhizoma Coptidis and Cortex Phellodendri, with 95% alcohol reflux 5 times, merge alcohol extract, placement is spent the night, and centrifugal filtration keeps filtrate, and water washing and precipitating is clear and bright to water lotion, precipitates standby;
Filtrate and water lotion are merged, regulate pH to pH=4 with concentrated hydrochloric acid, standing over night is separated out precipitation, and water washing and precipitating is to neutral;
Merge the gained precipitation 2 times, 80 ℃ of vacuum dryings are pulverized, and cross 200 mesh sieves, and obtaining total alkaloids is Rhizoma Coptidis and Cortex Phellodendri effective site.
Embodiment 2: the preparation of Chinese medicine extract of the present invention:
Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae, Radix Et Rhizoma Rhei effective site and the Rhizoma Coptidis, the Cortex Phellodendri effective site mixing that make among the embodiment 1 are promptly got Chinese medicine extract of the present invention.
Embodiment 3: the preparation of Chinese medicine extract capsule of the present invention:
Get the Chinese medicine extract of preparation among the embodiment 1, the snap fit capsule of packing into makes medicine capsule of the present invention.
Embodiment 4: Chinese medicine extract granule of the present invention must prepare:
Get the Chinese medicine extract of preparation among the embodiment 1, be ground into fine powder, add ethanol and make adhesive, add starch and make implant, be pressed into granule.
Embodiment 5: the preparation of Chinese medicinal tablet active component of the present invention:
Get Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae, Radix Et Rhizoma Rhei effective site and Rhizoma Coptidis, the Cortex Phellodendri effective site that embodiment 1 makes and evenly promptly get active component of the present invention by 3.5: 1 mixed.
Embodiment 6: the preparation of Chinese medicinal tablet of the present invention:
Embodiment 6-1
(1) granulates: take by weighing active component and the carboxymethyl starch sodium that embodiment 5 makes in 3: 1 ratio of mass ratio according to active component and carboxymethyl starch sodium, and carboxymethyl starch sodium is divided into two fens, portion is that 1/3 amount is standby, another part is that 2/3 amount joins in the active component, with concentration is that 70% ethanol is that wetting agent is granulated, and the granule that makes is dry down in 60 ℃;
(2) tabletting: get dried particles, add the carboxymethyl starch sodium of 1/3 standby amount and the magnesium stearate of tablet total weight amount 0.3%, depress to tablet with 3kN pressure.
Embodiment 6-2
(1) granulates: take by weighing active component and carboxymethyl starch sodium in 3: 1 ratio of mass ratio according to active component and carboxymethyl starch sodium, and carboxymethyl starch sodium is divided into two fens, portion is that 1/3 amount is standby, another part is that 2/3 amount joins in the active component, with concentration is that 75% ethanol is that wetting agent is granulated, and the granule that makes is dry down in 60 ℃;
(2) tabletting: get dried particles, add the carboxymethyl starch sodium of 1/3 standby amount and the magnesium stearate of tablet total weight amount 0.3%, depress to tablet with 5kN pressure.
Embodiment 6-3
(1) granulates: take by weighing active component and carboxymethyl starch sodium in 3: 1 ratio of mass ratio according to active component and carboxymethyl starch sodium, and carboxymethyl starch sodium is divided into two fens, portion is that 1/3 amount is standby, another part is that 2/3 amount joins in the active component, with concentration is that 80% ethanol is that wetting agent is granulated, and the granule that makes is dry down in 60 ℃;
(2) tabletting: get dried particles, add the carboxymethyl starch sodium of 1/3 standby amount and the magnesium stearate of tablet total weight amount 0.3%, depress to tablet with 7kN pressure.
The above only is an illustrative, but not is restricted person.Anyly do not break away from spirit of the present invention and category, and, all should be contained in the claim of the present invention its equivalent modifications of carrying out or change.
Claims (8)
1. Chinese medicine extract for the treatment of hepatitis, made by crude drug Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae, Radix Et Rhizoma Rhei, Rhizoma Coptidis and Cortex Phellodendri, it is characterized in that: it is made up of the effective site of Herba Artemisiae Scopariae 5.56kg, Fructus Gardeniae 3.70kg, Radix Scutellariae 1.85kg and the Radix Et Rhizoma Rhei 1.85kg of following weight proportion and the effective site of Rhizoma Coptidis 1.85kg and Cortex Phellodendri 1.85kg; The effective site of wherein said Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei makes by following step:
(1), get Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei decocting and boil 3 times, each 0.5～2 hour, merge decoction liquor, being evaporated to relative density is 1.05～1.10, standing over night, collecting precipitation and filtrate respectively after the centrifugal filtration,
(2), filtrate adds pretreated D101 macroporous adsorptive resins purification, with 50%～90% ethanol elution, collects pure washing liquid,
(3), with the precipitation in the step (1) with 6 times of amount alcohol refluxs of 95% 1 hour, filter, pure washing liquid in filtrate and the step (2) is merged, decompression recycling ethanol, being concentrated into 80 ℃ of relative densities of surveying down is 1.25～1.30, at 70 ℃ of vacuum dryings, pulverized 100 mesh sieves, obtain the effective site of Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei;
The effective site of described Rhizoma Coptidis and Cortex Phellodendri makes by following step:
(1), get Rhizoma Coptidis, Cortex Phellodendri with 85%～95% alcohol reflux 1～5 time, merge alcohol extract, spend the night, get supernatant, reclaim under reduced pressure is spent the night to there not being the alcohol flavor, centrifugal filtration, filtrate and precipitation,
(2), water washing and precipitating to water lotion clarifies, and precipitates standby.
(3), the water lotion in filtrate in the step (1) and the step (2) is merged, transfer pH value to 1～4 with concentrated hydrochloric acid, spend the night, separate out precipitation, water washing and precipitating is to neutral.
(4), with in the step (2) with step (3) in washing after precipitate merge, 60～80 ℃ of vacuum dryings are pulverized, and cross 50～200 mesh sieves, obtain the effective site of Rhizoma Coptidis and Cortex Phellodendri.
2. the preparation method of the Chinese medicine extract of the described treatment hepatitis of claim 1, it comprises following step:
(1), take by weighing Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae, Radix Et Rhizoma Rhei, Rhizoma Coptidis and the Cortex Phellodendri of described weight proportion, standby;
(2), get Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei decocting and boil 3 times, each 0.5～2 hour, merge decoction liquor, collecting precipitation and filtrate respectively after the centrifugal filtration;
(3), filtrate adds pretreated D101 macroporous adsorptive resins purification, with 50%～90% ethanol elution, collects pure washing liquid;
(4), with the precipitation in the step (1) with 6 times of amount alcohol refluxs of 95% 1 hour, filter, pure washing liquid in filtrate and the step (2) is merged, decompression recycling ethanol, being concentrated into 80 ℃ of relative densities of surveying down is 1.25～1.30, at 70 ℃ of vacuum dryings, pulverized 100 mesh sieves, obtain the effective site of Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei;
(5), get Rhizoma Coptidis, Cortex Phellodendri with 85%～95% alcohol reflux 1～5 time, merge alcohol extract, spend the night, get supernatant, reclaim under reduced pressure is spent the night to there not being the alcohol flavor, centrifugal filtration, filtrate and precipitation;
(6), water washing and precipitating to water lotion clarifies, and precipitates standby;
(7), the water lotion in filtrate in the step (1) and the step (2) is merged, transfer pH value to 1～4 with concentrated hydrochloric acid, spend the night, separate out precipitation, water washing and precipitating is to neutral;
(8), with in the step (2) with step (3) in washing after precipitate merge, 60 ℃～80 ℃ vacuum dryings are pulverized, and cross 50～200 mesh sieves, obtain the effective site of Rhizoma Coptidis and Cortex Phellodendri;
(9), get the effective site of Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei that step (4) obtains and Rhizoma Coptidis that step (8) obtains and the effective site of Cortex Phellodendri, mix homogeneously promptly gets Chinese medicine extract.
3. preparation method according to claim 2, it is characterized in that: the D101 macroporous adsorbent resin blade diameter length ratio in the described step (3) is 1: 5～1: 9, medical material and resin are 1: 2～1: 4, flow velocity is 2～3 times of resin bed volumes per hour, cross the post liquid stream to the greatest extent after, washing, be negative to judging terminal point with the Molish reaction, continue with 50%～90% ethanol elution, add to the eluent till the strong aqua ammonia permanent red, collect pure washing liquid.
4. Chinese medicinal tablet for the treatment of hepatitis, it is characterized in that: it comprises active component and pharmaceutically acceptable tablet adjuvant commonly used, and the effective site that effective site that wherein said active component is made by Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and the Radix Et Rhizoma Rhei of the described weight proportion of claim 1 and Rhizoma Coptidis and Cortex Phellodendri make is formed by 3.5: 1 weight ratio; The effective site of described Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei makes by following step:
(1), get Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei decocting and boil 3 times, each 0.5～2 hour, merge decoction liquor, being evaporated to relative density is 1.05～1.10, standing over night, collecting precipitation and filtrate respectively after the centrifugal filtration,
(2), filtrate adds pretreated D101 macroporous adsorptive resins purification, with 50%～90% ethanol elution, collects washing liquid.
(3), with the precipitation in the step (1) with 6 times of amount alcohol refluxs of 95% 1 hour, filter, pure washing liquid in filtrate and the step (2) is merged, decompression recycling ethanol, being concentrated into 80 ℃ of relative densities of surveying down is 1.25～1.30, at 70 ℃ of vacuum dryings, pulverized 100 mesh sieves, obtain the effective site of Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei;
The effective site of described Rhizoma Coptidis and Cortex Phellodendri makes by following step:
(1), get Rhizoma Coptidis, Cortex Phellodendri with 85%～95 alcohol reflux 1～5 time, merge alcohol extract, spend the night, get supernatant, reclaim under reduced pressure is spent the night to there not being the alcohol flavor, centrifugal filtration, filtrate and precipitation,
(2), water washing and precipitating to water lotion clarifies, precipitate standby,
(3), the water lotion in filtrate in the step (1) and the step (2) is merged, transfer pH value to 1～4 with concentrated hydrochloric acid, spend the night, separate out precipitation, water washing and precipitating is to neutral,
(4), with in the step (2) with step (3) in washing after precipitate merge, 60 ℃～80 ℃ vacuum dryings are pulverized, and cross 50～200 mesh sieves, obtain the effective site of Rhizoma Coptidis and Cortex Phellodendri.
5. the Chinese medicinal tablet of treatment hepatitis according to claim 4 is characterized in that: described tablet adjuvant commonly used is a carboxymethyl starch sodium, and the mass ratio of itself and active component is 1: 3.
6. the preparation method of the Chinese medicinal tablet of claim 4 or 5 described treatment hepatitis, it comprises following step:
(1), take by weighing Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae, Radix Et Rhizoma Rhei, Rhizoma Coptidis and the Cortex Phellodendri of described weight proportion, standby;
(2), get Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei decocting and boil 3 times, each 0.5～2 hour, merge decoction liquor, collecting precipitation and filtrate respectively after the centrifugal filtration;
(3), filtrate adds pretreated D101 macroporous adsorptive resins purification, with 50%～90% ethanol elution, collects pure washing liquid;
(4), with the precipitation in the step (1) with 6 times of amount alcohol refluxs of 95% 1 hour, filter, pure washing liquid in filtrate and the step (2) is merged, decompression recycling ethanol, being concentrated into 80 ℃ of relative densities of surveying down is 1.25～1.30, at 70 ℃ of vacuum dryings, pulverized 100 mesh sieves, obtain the effective site of Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei;
(5), get Rhizoma Coptidis, Cortex Phellodendri with 85%～95% alcohol reflux 1～5 time, merge alcohol extract, spend the night, get supernatant, reclaim under reduced pressure is spent the night to there not being the alcohol flavor, centrifugal filtration, filtrate and precipitation;
(6), water washing and precipitating is clear and bright to water lotion, precipitates standby;
(7), the water lotion in filtrate in the step (1) and the step (2) is merged, transfer pH value to 1～4 with concentrated hydrochloric acid, spend the night, separate out precipitation, water washing and precipitating is to neutral;
(8), with in the step (2) with step (3) in washing after precipitate merge, 60 ℃～80 ℃ vacuum dryings are pulverized, and cross 50～200 mesh sieves, obtain the effective site of Rhizoma Coptidis and Cortex Phellodendri;
(9), get the effective site of Herba Artemisiae Scopariae, Fructus Gardeniae, Radix Scutellariae and Radix Et Rhizoma Rhei that step (4) obtains and Rhizoma Coptidis that step (8) obtains and the effective site of Cortex Phellodendri, after mixing by 3.5: 1, add tablet adjuvant commonly used, make tablet.
7. preparation method according to claim 6 is characterized in that: the decoction process of described step (2) adds Herba Artemisiae Scopariae and Fructus Gardeniae earlier when decocting for the first time, boil the back and add Radix Scutellariae, adds Radix Et Rhizoma Rhei after little the boiling again, and begins to calculate decocting time.
8. preparation method according to claim 6, it is characterized in that: the D101 macroporous adsorbent resin blade diameter length ratio in the described step (3) is 1: 5～1: 9, medical material and resin are 1: 2～1: 4, flow velocity is 2～3 times of resin bed volumes per hour, cross the post liquid stream to the greatest extent after, washing, be negative to judging terminal point with the Molish reaction, continue with 50%～90% ethanol elution, add to the eluent till the strong aqua ammonia permanent red, collect pure washing liquid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100029796A CN101002849B (en) | 2007-01-30 | 2007-01-30 | Traditional Chinese medicine extractive or tablet for treating hepatitis, and its preparing method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100029796A CN101002849B (en) | 2007-01-30 | 2007-01-30 | Traditional Chinese medicine extractive or tablet for treating hepatitis, and its preparing method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101002849A CN101002849A (en) | 2007-07-25 |
| CN101002849B true CN101002849B (en) | 2011-01-26 |
Family
ID=38702375
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007100029796A Expired - Fee Related CN101002849B (en) | 2007-01-30 | 2007-01-30 | Traditional Chinese medicine extractive or tablet for treating hepatitis, and its preparing method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101002849B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101757121B (en) * | 2009-08-19 | 2011-07-20 | 李俊芳 | Internally and externally administered Chinese medicinal composition for curing gallbladder polyps |
| CN102631430B (en) * | 2012-04-19 | 2013-11-13 | 上海中医药大学 | Traditional Chinese medicine composition for resisting hepatic fibrosis, as well as preparation method and application thereof |
| CN102698114B (en) * | 2012-06-28 | 2013-12-04 | 陈慧婷 | Chinese medicinal composition comprising Radix Scutellariae for treating tympanites, and preparation method thereof |
| CN103316132B (en) * | 2013-06-03 | 2015-10-21 | 山西农业大学 | A kind of compound medicine preventing and treating liver and gall syndrome of fish |
| CN109453108B (en) * | 2018-12-06 | 2021-11-26 | 漳州卫生职业学院 | Coptis chinensis detoxifying nose-used temperature-sensitive gel and preparation method thereof |
| CN113546147A (en) * | 2021-09-04 | 2021-10-26 | 河南中医药大学 | Traditional Chinese medicine enema for treating hepatic encephalopathy |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1513486A (en) * | 2002-12-31 | 2004-07-21 | 财团法人工业技术研究院 | Liver-care composition and its prepn. method |
| CN1706442A (en) * | 2005-05-30 | 2005-12-14 | 许建文 | Prepn process of medicine for treating icteric diseases and oral liquid containing the medicine |
| CN1739650A (en) * | 2004-08-24 | 2006-03-01 | 芜湖绿叶制药有限公司 | Cappillary artemisia-cape jasmine-rhubarb soft capsule and its prepn process |
-
2007
- 2007-01-30 CN CN2007100029796A patent/CN101002849B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1513486A (en) * | 2002-12-31 | 2004-07-21 | 财团法人工业技术研究院 | Liver-care composition and its prepn. method |
| CN1739650A (en) * | 2004-08-24 | 2006-03-01 | 芜湖绿叶制药有限公司 | Cappillary artemisia-cape jasmine-rhubarb soft capsule and its prepn process |
| CN1706442A (en) * | 2005-05-30 | 2005-12-14 | 许建文 | Prepn process of medicine for treating icteric diseases and oral liquid containing the medicine |
Non-Patent Citations (6)
| Title |
|---|
| 刘太华等.大孔吸附树脂富集茵陈蒿汤中蒿属香豆素的工艺研究.中草药34 7.2003,34(7),603-604. |
| 刘太华等.大孔吸附树脂富集茵陈蒿汤中蒿属香豆素的工艺研究.中草药34 7.2003,34(7),603-604. * |
| 慕永平等.茵陈蒿汤的发展及现代研究.中国实验方剂学杂志12 2.2006,12(2),67-70. |
| 慕永平等.茵陈蒿汤的发展及现代研究.中国实验方剂学杂志12 2.2006,12(2),67-70. * |
| 王立福等.中西医结合治疗慢性重度乙型黄疸型肝炎69例.中医药信息23 4.2006,23(4),40-41. |
| 王立福等.中西医结合治疗慢性重度乙型黄疸型肝炎69例.中医药信息23 4.2006,23(4),40-41. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101002849A (en) | 2007-07-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101002849B (en) | Traditional Chinese medicine extractive or tablet for treating hepatitis, and its preparing method | |
| CN101396544B (en) | Traditional Chinese medicine composition capable of ventilating the lung and relieving asthma and preparation method thereof | |
| CN101332284B (en) | Chinese traditional medicine composition for treating ulcerative colitis and its preparation method | |
| CN102210737B (en) | Sweet potato leaf extract and preparation method and use thereof | |
| CN100584348C (en) | Anti-hepatitis medical combination | |
| CN1985864B (en) | Medicine composition prepared mainly from glycyrrhizic acid or its salt, ginseng and glossy ganoderma | |
| CN1977885B (en) | Antihepatitis medicinal composition | |
| CN1985873B (en) | Medicine composition of glycyrrhizic acid or its salt, ginseng and astragalus root | |
| CN102885928A (en) | Chinese medicinal injection for treating jaundice and preparation method thereof | |
| CN102178786A (en) | Medicament combination for treating diabetic peripheral neuropathy as well as preparation method and application thereof | |
| CN104352531B (en) | Application of the swinging seat nematode grass in hepatic and health products is prepared | |
| CN102188461B (en) | Application of screwtree root extractive in the preparation of anti-HBV medicament | |
| CN103040857B (en) | Application of phyllanthus urinaria polysaccharide component in preparing drug for resisting hepatitis B virus | |
| CN103143012B (en) | Traditional Chinese medicine composition and applications in preparation of porcine pseudorabies vaccine immunopotentiator | |
| CN102362993B (en) | Chinese medicinal preparation with effects of protecting intestines and removing toxic materials, and preparation method thereof | |
| CN100388928C (en) | Hepatitis virus resistant Chinese medicinal formulation and method for preparing same | |
| CN102940621B (en) | Application of methyl ferulic acid in preparation of medicine for preventing and curing hepatic fibrosis | |
| CN105943681A (en) | Semen litchi steroid saponin extract having function of improving insulin resistance | |
| CN100421700C (en) | Xianxiang medicine for treating hepatitis B and its production method | |
| CN1970001B (en) | Pharmaceutical composition comprising kurarinone, magnolia vine fruit and ginseng for treating hepatitis | |
| CN1969937B (en) | Pharmaceutical composition for treating hepatitis | |
| CN1961894B (en) | A novel compound pharmaceutical composition, preparation method and use thereof | |
| CN103610798A (en) | Active site of oldenlandia diffusa as well as preparation method and application thereof | |
| CN105017376B (en) | A kind of Herba Sarcopyramis nepalensis steroidal saponin and application thereof | |
| CN104173681B (en) | Traditional Chinese herbal medicine composition for treating castration-resistant prostate cancer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110126 Termination date: 20180130 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |