CN101940642A - Chinese medicinal composition and application thereof - Google Patents
Chinese medicinal composition and application thereof Download PDFInfo
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- CN101940642A CN101940642A CN2010105015607A CN201010501560A CN101940642A CN 101940642 A CN101940642 A CN 101940642A CN 2010105015607 A CN2010105015607 A CN 2010105015607A CN 201010501560 A CN201010501560 A CN 201010501560A CN 101940642 A CN101940642 A CN 101940642A
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Abstract
The invention discloses a Chinese medicinal composition. The Chinese medicinal composition comprises the following components: a capillary wormwood herb extract, a cape jasmine fruit extract, a rhubarb extract I and a rhubarb extract II, and the weight (dry weight) ratio of the components is 1:0.2:0.2:0.2-1:10:15:15. The Chinese medicinal composition can effectively improve the indexes of liver function and relieve the extent of liver fibrosis, and has better curative effect compared with that of the compatibility prescriptions of raw medicinal materials; moreover, the components of the Chinese medicinal composition are definite, and the quality is easy to control.
Description
Technical field
The present invention is specifically related to a kind of Chinese medicine composition and application thereof.
Background technology
Viral hepatitis is a kind of infectious disease, and domestic hepatitis virus carrier is about 1.2 hundred million, and hepatitis reaches more than 3,000 ten thousand.The ratio that hepatitis develops into hepatic fibrosis, liver cirrhosis and even hepatocarcinoma is very high, has had a strong impact on patient's life-span and life quality.In addition, the sickness rate of primary biliary cirrhosis and drug-induced liver disease has ever-increasing trend.This shows that the medicine of exploitation treatment liver and gall diseases has great social significance.
The Herba Artemisiae Scopariae Decoction beginning is stated from the works treatise on Febrile Diseases of Han dynasty Zhang Zhongjing: " this is that heat is got over, and can not turn to be yellow also for YANG MING syndrome, fever and sweating person; Sweating over the head only, body does not have heat, agent neck and going back, dysuria, the person that draws the slurry yearningly, this is an accumulation of heat in the interior, body must turn to be yellow, the Herba Artemisiae Scopariae Decoction master it.Six liang of Herba Artemisiae Scopariae, 14 pieces of Fructus Gardeniaes, two liang of Radix Et Rhizoma Rhei.Right three flavors, a pipeful of with water, boil Herba Artemisiae Scopariae earlier, subtract six liters, interior two flavors boil and get three liters, go dregs, divide temperature three clothes, urine is when sharp, urine is as the Fructus Gleditsia sap-shape, color is just red, one night abdominal distention reduced, Huang goes also from urine." Herba Artemisiae Scopariae is the special medicine of the subcutaneous ulcer of harnessing the Yellow River in the side, it is hot and suffocating to let out liver and gall clearly, and the energy dampness removing is monarch drug with the jaundice eliminating subcutaneous ulcer; The Fructus Gardeniae clearing away damp-heat from SANJIAO is damp and hot, is accessory drugs; Radix Et Rhizoma Rhei falls lets out stagnant heat, is adjuvant.A large amount of statistics show that in the treatment for icterohepatitis, Herba Artemisiae Scopariae Decoction shortens the course of disease for reducing icteric index, and mitigation symptoms and recovery liver function all have remarkable result.
Herba Artemisiae Scopariae is the dry aerial parts of feverfew BINHAO Artemisia scoparia Waldst.et Kit. or Herba Artemisiae Scopariae Artemisia capillaries Thunb..Gather during the high 6～10cm of seedling in spring or autumn alabastrum grow up to spending when just opening and tap, remove impurity and old footpath, dry.The habit of gathering spring claims " capillary wormwood ", the title of tapping autumn " flower Herba Artemisiae Scopariae " (Chinese Pharmacopoeia 2010 editions, 223 pages).Herba Artemisiae Scopariae hardship, suffering, be slightly cold, return spleen, stomach, liver, gallbladder meridian, clearing away heat-damp and promoting diuresis, promoting the function of the gallbladder to alleviate jaundice.Mainly contain chemical constituents such as volatile oil, organic acid, flavone, coumarin, chromogen ketone in the Herba Artemisiae Scopariae, wherein the organic acid chemical compound is seedling and the highest effective ingredient of the period content of putting forth, and representational organic acid has chlorogenic acid, isochlorogenic acid A/B/C, neochlorogenic acid and isomer thereof etc.Contain more organic acid (chlorogenic acid and Resina Ferulae acyl quinic acid class), flavone compound (Characterization of phenolic compounds inthe Chinese herbal drug Artemisia annua by liquid chromatography coupled toelectrospray ionization mass spectrometry.JOURNAL OF PHARMACEUTICALAND BIOMEDICAL ANALYSIS volume: 47 phases: 3 pages: 516-525), contain the higher coumarin of content, chromone constituents in the alabastrum of flower Herba Artemisiae Scopariae in the flower Herba Artemisiae Scopariae.No matter be capillary wormwood or colored Herba Artemisiae Scopariae, its chlorogenic acid chemical compound can effectively suppress lipid peroxide to hepatocellular destruction, has hepatoprotective effect.
Fructus Gardeniae is the dry mature fruit of Maguireothamnus speciosus SHANZHIZI Gardenia jasminoides Ellis, and hardship is cold.GUIXIN, lung, tri-jiao channel.The pathogenic fire purging relieving restlessness is arranged, clearing away heat and promoting diuresis, the effect of removing pathogenic heat from blood and toxic substance from the body.Mainly contain chemical compounds such as iridoid glycosides, crocin class, chlorogenic acid, volatile oil, polysaccharide in the Fructus Gardeniae, contain tens kinds of iridoid glycoside compounds in the present known Fructus Gardeniae, comprise jasminoidin, Gardenoside, Fructus Gardeniae thuja acid, geniposide etc.Jasminoidin is a kind of important iridoids glycosides in the Fructus Gardeniae, has very high medical value.It has heat clearing away and separates effect tired, cooling blood for hemostasis, can be used as antibacterial and formula for clearing heat and removing toxicity; Jasminoidin also has the effect of cholagogue and eliminating inflammation, is used for acute and chronic cholecystitis, cholelithiasis and biliary tract infection etc. clinically.
Radix Et Rhizoma Rhei is the dry root and rhizome of polygonum rheum palmatum Rheum palmatum L., Rheum tanguticum Rheumtanguticum Maxim.ex Balf. or Rheum officinale Rheum officinale Baill..Radix Et Rhizoma Rhei hardship, cold.Return spleen, stomach, large intestine, liver, pericardium channel.Purging heat and dredging bowels, removing pathogenic heat from blood and toxic substance from the body, eliminating blood stasis and inducing menstruation.The processed product of Radix Et Rhizoma Rhei has polytype, and as Radix et Rhizoma Rhei (stir-fried with wine), Radix Et Rhizoma Rhei, charred Radix Et Rhizoma Rhei etc., Radix et Rhizoma Rhei (stir-fried with wine) is apt to clear part of the body cavity above the diaphragm housing the heart and lungs noxious heat in the blood system; Radix Et Rhizoma Rhei rushes down exerts oneself to delay eliminating fire and detoxication; The hemostasis of charred Radix Et Rhizoma Rhei cooling blood and removing stasis.The chemical constituent that Radix Et Rhizoma Rhei contained is of a great variety, present known have tannin class, anthraquinone derivative class, stilbene class, phenyl propyl ketone glycoside, naphthalene derivatives class, chromogen ketone etc.Tanning and precursor thereof account for 10%～30% of the total composition of Radix Et Rhizoma Rhei, belong to have more than 30 of hydrolysis-type tannin, and what belong to the condensed type tannin has 37.Anthraquinone derivative accounts for 3%～5% of the total composition of Radix Et Rhizoma Rhei, major part be glucose in conjunction with glycosides, free aglycon has chrysophanic acid, emodin, aloe-emodin, chrysophanol, physcione etc.The dianthrone class has palm leaf dianthrone A, B, C, reidin A, B, C and Sennoside A, B, C, D, etc.In addition, Radix Et Rhizoma Rhei also contains saccharide, organic acid, inorganic matter, free amino acid and protein, volatile compound etc.After concocting, the Senna fruit glycoside content with strong discharge function sharply descends, so Radix et Rhizoma Rhei (processed) rushes down than Radix Et Rhizoma Rhei and exerts oneself to delay.Radix Et Rhizoma Rhei have protect the liver, choleretic effect, can obviously reduce the SGPT activity, recover hepatocellular normal function, promote hepatocellular regeneration regulating liver-QI cell RNA synthetic; Stimulate human body to produce interferon; Strengthen the dog gallbladder and shrink, lax oddi's sphincter; Blood capillary in the mediation liver, thus the recovery of promotion liver function stops light-duty hepatitis to transform to hepatitis gravis, reduces complication, reduces mortality rate.
Tradition Herba Artemisiae Scopariae Decoction decoction is to make up a prescription, and adds water and fries in shallow oil altogether and get, and this method exists dose big, inconvenience, many shortcomings such as quality is uncontrollable.Chinese medicine compound is a complex system, and the material that plays the curative effect effect is a class or the activated chemical compound of multiclass normally, and they play primary and secondary, many target spots, the collaborative therapeutic effect of organic whole.With chemical compounds dissimilar in the compound recipe, adopt modern extraction, separation method, it is clear and definite relatively to be made into composition, the extract (being active component) that active constituent content is controlled, be mixed with tcm components by component compatibility then, reaching the effect of attenuation synergistic, reduce the dose of medicine simultaneously, is a kind of new way of exploitation modern Chinese medicine preparation.
Summary of the invention
Technical problem to be solved by this invention is to have defectives such as dose is big, inconvenient, quality is uncontrollable in order to overcome traditional Herba Artemisiae Scopariae Decoction decoction, and a kind of Chinese medicine composition and application thereof are provided.Chinese medicine composition of the present invention can effectively improve liver function index, alleviates the degree of hepatic fibrosis that is caused by cholestasis, and curative effect is better than the curative effect of crude drug compatibility side, and drug component is clear, and quality is controlled easily.
The present invention relates to a kind of Chinese medicine composition, it comprises following ingredients: Herba Artemisiae Scopariae extract, Fructus Gardeniae extract, Radix Et Rhizoma Rhei extract I and Radix Et Rhizoma Rhei extract II; Wherein, the ratio of weight and number of described Herba Artemisiae Scopariae extract, Fructus Gardeniae extract, Radix Et Rhizoma Rhei extract I and Radix Et Rhizoma Rhei extract II is 1: 0.2: 0.2: 0.2～1: 10: 15: 15, preferable was 1: 0.2: 0.2: 0.2～1: 5: 5: 5; Better is 1: 1: 1: 1～1: 3: 10: 10; Above ratio is the ratio of weight and number of each extract dry weight;
In the described Herba Artemisiae Scopariae extract, the organic acid chemical compound accounts for the quality percentage composition of Herba Artemisiae Scopariae extract dry weight greater than 8%, and is preferred 10%～15%, and wherein, chlorogenic acid accounts for the quality percentage composition of Herba Artemisiae Scopariae extract dry weight more than or equal to 3%, and better is 4.7%～6%; In the described Fructus Gardeniae extract, jasminoidin accounts for the quality percentage composition of Fructus Gardeniae extract dry weight more than or equal to 40%, and preferred 55%～60%; Among the described Radix Et Rhizoma Rhei extract I, rhubarb polyoses accounts for the quality percentage composition of Radix Et Rhizoma Rhei extract I dry weight greater than 40%, and preferred 62%～65%; Among the described Radix Et Rhizoma Rhei extract II, rhatannin accounts for the quality percentage composition of Radix Et Rhizoma Rhei extract II dry weight more than or equal to 8%, and is preferred 10%～12%, wherein, gallic acid accounts for the quality percentage composition of Radix Et Rhizoma Rhei extract II dry weight more than or equal to 1%, is preferably more than or equals 2%.
Among the present invention, what described Herba Artemisiae Scopariae was preferable is dry aerial parts or its processed product of feverfew BINHAO or Herba Artemisiae Scopariae; What described Fructus Gardeniae was preferable is dry mature fruit or its processed product of Maguireothamnus speciosus Fructus Gardeniae; What described Radix Et Rhizoma Rhei was preferable is dry root, rhizome or its processed product of polygonum rheum palmatum, Rheum tanguticum or Rheum officinale.
Among the present invention, the raw material of preparation Herba Artemisiae Scopariae extract, Fructus Gardeniae extract and two kinds of Radix Et Rhizoma Rhei extracts, what promptly the ratio of weight and number of Herba Artemisiae Scopariae, Fructus Gardeniae and Radix Et Rhizoma Rhei was preferable is 1: (0.1-2): (0.1-2).
Among the present invention, described Herba Artemisiae Scopariae extract can make according to the method among the Chinese patent CN101580447A.Described Fructus Gardeniae extract can make by Chinese invention patent CN101012248A (application number is 200710037097.3) method.
Described Herba Artemisiae Scopariae extract can be made by following method:
(1) be that 0%～30% ethanol water joins in the extraction equipment with capillary wormwood medical material and percent by volume, heating and refluxing extraction 0.5～2 hour, ethanol water is 10～25 times of capillary wormwood quality of medicinal material, extract 1～3 time, make the capillary wormwood extracting solution after filtration, be evaporated to 1～3 times of capillary wormwood medical material volume;
(2) add ethanol water in the capillary wormwood extracting solution that step (1) makes, the percent by volume of ethanol water is 60%～80%, places 10～14 hours, centrifugal, filtration obtains clear liquid, is evaporated to 1～3 times of capillary wormwood medical material volume, makes the capillary wormwood concentrated solution;
(3) the capillary wormwood concentrated solution that step (2) is made is pH to 2～5 of the hydrochloric acid solution adjusting capillary wormwood concentrated solution of 0.1～2mol/L with concentration, filters, and makes capillary wormwood filtrate;
(4) the capillary wormwood filtrate that step (3) is made, through absorption with macroporous adsorbent resin, earlier with the water elution impurity of 2～3 times of column volumes and discard cleanout fluid, 3～6 times of column volumes of reuse, percent by volume are 40～60% ethanol water eluting, behind the eluent concentrate drying, make Herba Artemisiae Scopariae extract.
What wherein, described macroporous adsorbent resin was preferable is the styrene macroporous adsorbent resin.
Described Fructus Gardeniae extract can be made by following method:
(1 ') with the cape jasmine fruit coarse powder with the ethanol water of percent by volume 50%～80% heating and refluxing extraction 15～60 minutes in extraction equipment, ethanol water is 8～12 times of crude drug coarse powder quality, extract 1～3 time, obtain gardenia extract through centrifugal, filtration, be evaporated to 3～5 times of crude drug volume;
(2 ') will extract concentrated solution through absorption with macroporous adsorbent resin by the Fructus Gardeniae of step (1 ') gained, elder generation is 0～10% ethanol water eluting impurity with percent by volume, the back will promptly get Fructus Gardeniae extract with the ethanol water eluting of percent by volume 25%～35% behind the eluent concentrate drying.
What the use amount of used ethanol water was preferable when wherein, carrying out eluting in the step (2 ') is 3～6 times of column volumes.What described macroporous adsorbent resin was preferable is the styrene macroporous adsorbent resin.
Among the present invention, described Radix Et Rhizoma Rhei extract I can be made by following method:
(1 ") Radix Et Rhizoma Rhei water or ethanol water are carried out reflux, extract,, with extracting liquid filtering, get supernatant, promptly get Radix Et Rhizoma Rhei extract;
The Radix Et Rhizoma Rhei extract that (2 ") make step (1) is concentrated into volume mass with crude drug than being 0.5-3ml/g; add volumn concentration 95% ethanol water and reach 60%～80% (volumn concentration) to containing the alcohol amount; left standstill 8～24 hours; filter; with ethanol water washing precipitation; drying promptly gets Radix Et Rhizoma Rhei extract I.
Step (1 ") in, what the volumn concentration of described ethanol water was preferable is greater than 0, is less than or equal to 30%; What the time of described reflux, extract, was preferable is 10-60 minute, and what the number of times of extraction was preferable is 1-3 time.
Step (2 ") in, what the described temperature that leaves standstill was preferable is 2～10 ℃; What the volumn concentration of the ethanol water of washing precipitation was preferable is 50%～95%.
Among the present invention, described Radix Et Rhizoma Rhei extract II can be made by following method: will prepare the supernatant concentration that obtains after filtering in the step (2 ") of Radix Et Rhizoma Rhei extract I, drying gets final product.
Among the present invention, above-mentioned each composition is simply mixed, can make Chinese medicine composition of the present invention.
The invention further relates to above-mentioned Chinese medicine composition and treat and/or prevent application in the medicine of hepatic disease in preparation.What wherein, described hepatic disease was preferable is jaundice or the hepatic fibrosis that hepatitis, cholestasis cause.
As required, according to the knowledge of this area, Chinese medicine composition of the present invention can be made pharmaceutical composition with various typical additives (as diluent and excipient etc.) pharmaceutically.
Except that specified otherwise, raw material that the present invention relates to and reagent are all commercially available to be got.
Positive progressive effect of the present invention is:
1, Chinese medicine composition effective substance of the present invention is clear and definite, comprises Herba Artemisiae Scopariae organic acid extract, Fructus Gardeniae iridoid glycoside extract, rhubarb polyoses extract and rhatannin extract that proportion compatibility is clear and definite.
2, the preparation method process stabilizing of each composition in the Chinese medicine composition of the present invention, mass discrepancy is little between therefore prepared each batch of drug products that goes out.
3, the pharmaceutical formulation in the Chinese medicine composition of the present invention, the active component in Herba Artemisiae Scopariae organic acid extract, Fructus Gardeniae iridoid glycoside extract, rhubarb polyoses extract and the rhatannin extract can be surveyed, and therefore can effectively control the quality of drug products of the present invention.
4, the present invention induces hepatic fibrosis experiment, ANIT chmice acute hepatic injury experiment by rat common bile duct ligation hepatic fibrosis experiment, DMN, the result shows, adopt compatible in proportion the drug action of Herba Artemisiae Scopariae organic acid, Fructus Gardeniae iridoid glycoside, rhubarb polyoses and rhatannin with the best, can effectively improve liver function index, alleviate the degree of hepatic fibrosis that is caused by cholestasis, curative effect is better than the curative effect of crude drug compatibility side.
Description of drawings
When Figure 1A is wavelength 260nm, the HPLC finger printing of the Chinese medicine composition of the embodiment of the invention 1 preparation.
When Figure 1B is wavelength 330nm, the HPLC finger printing of the Chinese medicine composition of the embodiment of the invention 1 preparation.
Fig. 2 is the hepatic tissue pathology section (microphotograph) of the Chinese medicine composition drug particles of the embodiment of the invention 2 preparations to the treatment effect of N-nitrosodimethylamine (DMN) model.
The specific embodiment
Further specify the present invention with embodiment below, but the present invention is not limited.
The preparation of the medicinal tablet of embodiment 1, treatment hepatic disease
One, the preparation of Herba Artemisiae Scopariae extract
Herba Artemisiae Scopariae extract prepares with reference to Chinese invention patent 101580447A: accurately take by weighing capillary wormwood 1.8kg, adding the 32L volumn concentration is that 14% ethanol carries out reflux, extract,, and extraction time is 1h, collects extracting solution; And then adding 32L volumn concentration is that 14% ethanol carries out reflux, extract, in residue, and extraction time is 1h, collects extracting solution.Twice extracting solution is mixed, be cooled to room temperature, filter, get filtrate decompression and be concentrated into nothing alcohol flavor, obtain 3L extract extractum.Adding volumn concentration in extractum is 95% ethanol, is adjusted to that to contain the alcohol amount be 70%, and standing over night is filtered.Filtrate decompression is concentrated into does not have the alcohol flavor, is diluted with water to 2L, and the hydrochloric acid of reuse 0.3mol/L is regulated pH to 2.5.Centrifugal, get supernatant.
Adopt the good low pole macroporous adsorptive resins of pretreatment (filling with polystyrene or poly-divinylbenzene is that the model of skeleton is the resin extender (Shanghai China shake scientific ﹠ trading Co., Ltd.) of HZ-816) in resin column, the 2L Herba Artemisiae Scopariae extracting solution of above-mentioned acquisition is carried out separation and purification.The Herba Artemisiae Scopariae extracting solution with 1/4BV/h (1BV/h is promptly per hour by the flow of 1 times of bed volume) flow velocity upper prop absorption after, dash flow velocity flushing with 1BV/h except that resin column with the deionized water of 3 times of resin volumes, to remove water-solubility impurity such as polysaccharide, discard eluent; 3 times of volumn concentrations to the resin column bed volume of reuse are 50% the ethanol flow velocity eluting resin column with 1/2BV/h, collect eluent, obtain Herba Artemisiae Scopariae organic acid eluent.
In being evaporated to thick extractum below 60 ℃, 60 ℃ of vacuum dryings are to constant weight in vacuum drying oven then with above-mentioned eluent, and (organic acid content detects with HPLC and reaches 10%, and wherein chlorogenic acid contents is 4.7% to get capillary wormwood extract powder 28g.Chromatographic condition: with the octadecylsilane chemically bonded silica is filler; With volume fraction is that 0.1% acetic acid is mobile phase A, is Mobile phase B with the acetonitrile, gradient elution 0-30min, 5%-30%B; Flow velocity 0.8ml/min; Detect wavelength 324nm; 30 ℃ of column temperatures).
Two, the preparation of Fructus Gardeniae extract
Fructus Gardeniae extract prepares with reference to Chinese invention patent CN101012248A: accurately get dry cape jasmine fruit coarse powder 1.0kg, adding the 10L volumn concentration is 65% alcohol reflux, and extraction time 1h collects extracting solution.And then adding 10L volumn concentration is that 65% ethanol carries out reflux, extract, in residue, and extraction time is 1h, collects extracting solution.Extracting solution mixing with twice is cooled to room temperature, filters, and gets filtrate decompression and is concentrated into nothing alcohol flavor, and thin up extracting solution to volume/crude drug ratio is about 4: 1, and is centrifugal, gets supernatant.
Adopt the good low pole macroporous adsorptive resins of pretreatment (filling in resin column with polystyrene or poly-divinylbenzene is that the model of skeleton is the resin extender (Shanghai China shake scientific ﹠ trading Co., Ltd.) of HZ-816) that supernatant is carried out separation and purification.Supernatant with 5 times of bed volume washings, discards eluent with the absorption of 1/4BV/h flow velocity upper prop; Be the flow velocity eluting of 30% ethanol 1/2BV/h again with volumn concentration, collect eluent, obtain the Fructus Gardeniae iridoid glycoside eluent.
In being evaporated to thick extractum below 60 ℃, 60 ℃ of vacuum dryings are to constant weight in vacuum drying oven then with above-mentioned eluent, and (recording wherein with the HPLC method, the content of jasminoidin is 55% to get Fructus Gardeniae extract powder 58g.Chromatographic condition: with the octadecylsilane chemically bonded silica is filler; With volume fraction is 75% methanol aqueous solution mobile phase, flow velocity 0.7ml/min; Detect wavelength 238nm; 30 ℃ of column temperatures).
Three, the preparation of Radix Et Rhizoma Rhei extract
Accurately take by weighing Radix et Rhizoma Rhei (processed with wine) decoction pieces 600g, add the 6L deionized water and carry out reflux, extract,, extraction time is 15min, collects extracting solution; And then adding 4.2L deionized water carries out reflux, extract, in residue, and extraction time is 20min, collects extracting solution.Twice extracting solution is mixed, be cooled to room temperature, filter, get filtrate decompression and be concentrated into volume/crude drug ratio and be about 2: 1, adding percentage by volume is that 95% ethanol reaches 70% to containing the alcohol amount, and stand at low temperature is spent the night, and filters.
With above-mentioned filtrate in being evaporated to thick extractum below 60 ℃, 60 ℃ of vacuum dryings are to constant weight in vacuum drying oven then, and (recording wherein with visible spectrophotometry, the content of rhatannin is 10% to get the about 101g of Radix et Rhizoma Rhei (processed) tannin extract powder (Radix Et Rhizoma Rhei extract II).Recording wherein with the HPLC method, the content of gallic acid is 2%.Chromatographic condition: with the octadecylsilane chemically bonded silica is filler; Volume fraction is that 0.1% acetic acid is mobile phase A, is Mobile phase B with methanol, A: B=98: 2, and flow velocity 1.0ml/min; Detect wavelength 270nm; 30 ℃ of column temperatures).
With above-mentioned filtering residue percentage by volume is 95% washing with alcohol, 60 ℃ of vacuum dryings obtain Radix et Rhizoma Rhei (processed) polyoses extract 95g (Radix Et Rhizoma Rhei extract I) (with visible spectrophotometry---phenol sulfuric acid process record wherein the content of rhubarb polyoses is 62%) to constant weight in vacuum drying oven then.
Four, the preparation of the medicinal tablet of treatment hepatic disease
Get the Herba Artemisiae Scopariae extract powder 20g of above-mentioned steps one preparation, the Fructus Gardeniae extract powder 40g of above-mentioned steps two preparations, the Radix et Rhizoma Rhei (processed) tannin extract powder 65g and the Radix et Rhizoma Rhei (processed) polyoses extract powder 95g of above-mentioned steps three preparations, with four mix homogeneously, add the 44g lactose again, (weight that adds lactose be four extract powder weight sums 20%) and 44g microcrystalline Cellulose (weight of adding microcrystalline Cellulose be four extract weight sums 20%), behind the mix homogeneously, add an amount of 95% alcoholic solution and make soft material, after crossing 18～20 mesh sieves and making wet granular, under 60 ℃, drug particles is carried out drying, add mass fraction again and be 0.5% magnesium stearate 1g, behind the mixing, the obtain medical treatment medicinal tablet of hepatic disease of tabletting.
This medicinal tablet is carried out HPLC analyze, see Figure 1A and Figure 1B.
The preparation of embodiment 2 Chinese medicine compositions of the present invention
Herba Artemisiae Scopariae extract dried powder, Fructus Gardeniae extract dried powder, Radix et Rhizoma Rhei (processed) tannin extract dried powder, the Radix et Rhizoma Rhei (processed) polyoses extract dried powder got among the embodiment 1 are mixed in proportion, joining with normal saline is the component drug group solution of 4 variable concentrations, and the concrete concentration of each extract sees Table 1 in 4 medicine groups.
Table 1DMN rat liver fibrosis model experiment medicine is formed (g/ml)
| Group number | Herba Artemisiae Scopariae extract | Fructus Gardeniae extract | The Radix et Rhizoma Rhei (processed) polyoses extract | Radix et Rhizoma Rhei (processed) tannin extract |
| 1 | 0.0070 | 0.0057 | 0.0091 | 0.0350 |
| 2 | 0.0049 | 0.0078 | 0.0158 | 0.0108 |
| 3 | 0.0039 | 0.0099 | 0.0362 | 0.0390 |
| 4 | 0.0112 | 0.0121 | 0.0294 | 0.0148 |
The preparation of the medicinal tablet of embodiment 3, treatment hepatic disease
One, the preparation of Herba Artemisiae Scopariae extract
Herba Artemisiae Scopariae extract prepares with reference to Chinese invention patent 101580447A: accurately take by weighing capillary wormwood 1.8kg, adding the 20L volumn concentration is that 30% ethanol carries out reflux, extract,, and extraction time is 0.5h, collects extracting solution; And then adding 32L volumn concentration is that 5% ethanol carries out reflux, extract, in residue, and extraction time is 0.5h, collects extracting solution.Twice extracting solution is mixed, be cooled to room temperature, filter, get filtrate decompression and be concentrated into nothing alcohol flavor, obtain 3L extract extractum.Adding volumn concentration in extractum is 60% ethanol, is adjusted to that to contain the alcohol amount be 70%, and standing over night is filtered.Filtrate decompression is concentrated into does not have the alcohol flavor, is diluted with water to 2L, and the hydrochloric acid of reuse 2mol/L is regulated pH to 4.Centrifugal, get supernatant.
Adopt the good low pole macroporous adsorptive resins of pretreatment (filling with polystyrene or poly-divinylbenzene is that the model of skeleton is the resin extender (Shanghai China shake scientific ﹠ trading Co., Ltd.) of HZ-816) in resin column, the 2L Herba Artemisiae Scopariae extracting solution of above-mentioned acquisition is carried out separation and purification.The Herba Artemisiae Scopariae extracting solution with 1/4BV/h (1BV/h is promptly per hour by the flow of 1 times of bed volume) flow velocity upper prop absorption after, dash flow velocity flushing with 1BV/h except that resin column with the deionized water of 3 times of resin volumes, to remove water-solubility impurity such as polysaccharide, discard eluent; 6 times of volumn concentrations to the resin column bed volume of reuse are 50% the ethanol flow velocity eluting resin column with 1/2BV/h, collect eluent, obtain Herba Artemisiae Scopariae organic acid eluent.
In being evaporated to thick extractum below 60 ℃, 60 ℃ of vacuum dryings are to constant weight in vacuum drying oven then with above-mentioned eluent, and (organic acid content detects with HPLC and reaches 10%, and wherein chlorogenic acid contents is 4.7% to get capillary wormwood extract powder 28g.Chromatographic condition: with the octadecylsilane chemically bonded silica is filler; With volume fraction is that 0.1% acetic acid is mobile phase A, is Mobile phase B with the acetonitrile, gradient elution 0-30min, 5%-30%B; Flow velocity 0.8ml/min; Detect wavelength 324nm; 30 ℃ of column temperatures).
Two, the preparation of Fructus Gardeniae extract
Fructus Gardeniae extract prepares with reference to Chinese invention patent CN101012248A: accurately get dry cape jasmine fruit coarse powder 1.0kg, adding the 10L volumn concentration is 50% alcohol reflux, 15 minutes extraction times, collects extracting solution.And then adding 10L volumn concentration is that 80% ethanol carries out reflux, extract, in residue, and extraction time is 30 minutes, collects extracting solution.Extracting solution mixing with twice is cooled to room temperature, filters, and gets filtrate decompression and is concentrated into nothing alcohol flavor, and thin up extracting solution to volume/crude drug ratio is about 10: 1, and is centrifugal, gets supernatant.
Adopt the good low pole macroporous adsorptive resins of pretreatment (filling in resin column with polystyrene or poly-divinylbenzene is that the model of skeleton is the resin extender (Shanghai China shake scientific ﹠ trading Co., Ltd.) of HZ-816) that supernatant is carried out separation and purification.Supernatant is washed with the alcoholic acid aqueous solution (percent by volume 10%) of 5 times of bed volume with the absorption of 1/4BV/h flow velocity upper prop, discards eluent; Be the flow velocity eluting of 30% ethanol 1/2BV/h again with volumn concentration, collect eluent, obtain the Fructus Gardeniae iridoid glycoside eluent.
In being evaporated to thick extractum below 60 ℃, 60 ℃ of vacuum dryings are to constant weight in vacuum drying oven then with above-mentioned eluent, and (recording wherein with the HPLC method, the content of jasminoidin is 55% to get Fructus Gardeniae extract powder 58g.Chromatographic condition: with the octadecylsilane chemically bonded silica is filler; With volume fraction is 75% methanol aqueous solution mobile phase, flow velocity 0.7ml/min; Detect wavelength 238nm; 30 ℃ of column temperatures).
Three, the preparation of Radix Et Rhizoma Rhei extract
Accurately take by weighing Radix et Rhizoma Rhei (processed with wine) decoction pieces 600g, the ethanol water that adds 6L percent by volume 30% carries out reflux, extract,, and extraction time is 60min, collects extracting solution; And then adding 4.2L deionized water carries out reflux, extract, in residue, and extraction time is 30min, collects extracting solution.Twice extracting solution is mixed, be cooled to room temperature, filter, get filtrate decompression and be concentrated into volume/crude drug ratio and be about 2: 1, adding mass percent is that 95% ethanol reaches 70% to containing the alcohol amount, in 2～10 ℃ of standing over night, filters.
With above-mentioned filtrate in being evaporated to thick extractum below 60 ℃, 60 ℃ of vacuum dryings are to constant weight in vacuum drying oven then, and (recording wherein with visible spectrophotometry, the content of rhatannin is 10% to get the about 101g of Radix et Rhizoma Rhei (processed) tannin extract powder (Radix Et Rhizoma Rhei extract II).Recording wherein with the HPLC method, the content of gallic acid is 2%.Chromatographic condition: with the octadecylsilane chemically bonded silica is filler; Volume fraction is that 0.1% acetic acid is mobile phase A, is Mobile phase B with methanol, A: B=98: 2, and flow velocity 1.0ml/min; Detect wavelength 270nm; 30 ℃ of column temperatures).
With above-mentioned filtering residue mass percent 60% washing with alcohol, 60 ℃ of vacuum dryings obtain Radix et Rhizoma Rhei (processed) polyoses extract 95g (Radix Et Rhizoma Rhei extract I) (with visible spectrophotometry---phenol sulfuric acid process record wherein the content of rhubarb polyoses is 62%) to constant weight in vacuum drying oven then.
Four, the preparation of the medicinal tablet of treatment hepatic disease
Get the Herba Artemisiae Scopariae extract powder 20g of above-mentioned steps one preparation, the Fructus Gardeniae extract powder 40g of above-mentioned steps two preparations, the Radix et Rhizoma Rhei (processed) tannin extract powder 65g and the Radix et Rhizoma Rhei (processed) polyoses extract powder 95g of above-mentioned steps three preparations, with four mix homogeneously, add the 44g lactose again, (weight that adds lactose be four extract powder weight sums 20%) and 44g microcrystalline Cellulose (weight of adding microcrystalline Cellulose be four extract weight sums 20%), behind the mix homogeneously, add an amount of 95% alcoholic solution and make soft material, after crossing 18～20 mesh sieves and making wet granular, under 60 ℃, drug particles is carried out drying, add mass fraction again and be 0.5% magnesium stearate 1g, behind the mixing, the obtain medical treatment medicinal tablet of hepatic disease of tabletting.
The effect experiment of the drug extract of effect embodiment 1, treatment hepatic disease
Get 4 component drug solution that embodiment 2 makes.
Traditional decoction prepares by the following method: get Herba Artemisiae Scopariae 180g, add water 3.40L, soak 1h, decoct 40min, add Fructus Gardeniae (smashing to pieces) 100g again, Radix et Rhizoma Rhei (processed) 60g decocts 15min, filters extracting juice; Medicinal residues add water 2.4L, decoct 20min, filter extracting juice, merge 2 times filtrate, in being evaporated to thick extractum below 60 ℃, then in vacuum drying oven 60 ℃ of vacuum dryings to constant weight.Above-mentioned dried powder carries out effect experiment with the traditional decoction solution that normal saline is mixed with 0.13g/ml.
(180 ± 10g) are divided into 4 groups at random, are respectively normal group, model group, traditional decoction matched group, component drug group, 8 every group to get cleaning level male Wistar rat.Wherein, model group, traditional decoction matched group and component drug group are with N-nitrosodimethylamine (DMN) 10 μ l.kg
-1The dosage lumbar injection of rat body weight, every day 1 time, for three days on end, altogether 4 weeks of modeling, the normal rats intraperitoneal injection of saline is with the dosage normal saline.Traditional decoction matched group, component drug group are in the administration intervention of just dividing into groups in the 3rd week, according to rat body weight 10mlkg
-1Dosage is irritated the drug extract that stomach is treated hepatic disease, and once a day, administration is 14 days altogether; Model group rat and normal rats are irritated stomach according to the dosage of body weight and are given use tap water 10ml.kg
-1Give normal saline with volume.Make a collection of specimens after 4 weeks finished and do the index of correlation detection, carry out the pharmacodynamics controlled observation.
Drug treatment is pressed 2mlkg with 2% pentobarbital sodium after finishing
-1Dosage intraperitoneal injection of anesthesia rat, the posterior vena cava blood sampling, centrifugalize serum is got liver lobus sinister tissue, 10% formalin fixed, dehydration, transparent, paraffin embedding, slice thickness is 4 μ m, is used for collagen and immunohistochemical staining hepatic tissue.Detect serological index serum alanine aminotransferase (ALT), serum Aspartic Acid aminotransferase (AST), serum albumin (Alb), serum alkaline phosphatase (ALP), serum total bilirubin (TBiL), serum bile acid (TBA), serum gamma glutamyltransferase (GGT).Adopt the JamallShi method to measure hepatic tissue hydroxyproline (Hyp).Measurement data is write as average ± standard deviation
Form.Normal group and model group relatively reach model group and each treatment group is more all used the t check.Enumeration data is checked with Ridit.
Table 2 has been listed and has respectively been organized the influence of medicine to DMN hepatic fibrosis rats Serum ALT, AST, Alb, GGT, ALP, TBiL.Compare with normal group, DMN hepatic fibrosis rats Serum ALT, AST, GGT, ALP activity all obviously raise, and serum Alb content obviously reduces, TBiL content raises (P＜0.01).Compare with the model group rat, traditional decoction group and each component drug group rat blood serum ALT, AST, GGT, ALP activity all have decline in various degree, and serum Alb content obviously raises, TBiL content reduces (P＜0.01).Every index of component drug group 4 all is better than the traditional decoction group, component drug group 1 except that ALP, component drug group 2 except that ALT, other serological index all are better than the traditional decoction group, do not have significant difference between each component drug group and the traditional decoction group.
Organize the Hyp Determination on content to the results are shown in Table 3 to rat liver.With normal rat relatively, rat model hepatic tissue Hyp obviously raise (P＜0.01).With model group relatively, traditional decoction and component drug have in various degree reduction effect (P＜0.01) to liver tissues of rats Hyp.Component drug group 3 and component drug group 4 rat livers organize Hyp content slightly to be better than the traditional decoction group, component drug group 1 and component drug group 2 rat livers organize Hyp content close with the traditional decoction group, but do not have significant difference between each component drug group and the traditional decoction group.
Table 3 is respectively organized the influence of medicine to DMN hepatic fibrosis rats hepatic tissue Hyp content
| Group | Hyp (μ g/g wet liver) |
| Normal group | 244.096±44.936 |
| Model group | 573.491±103.013 |
| The traditional decoction group | 361.384±43.97 |
| Component drug group 1 | 362.582±42.537 |
| Component drug group 2 | 370.971±44.838 |
| Component drug group 3 | 339.514±51.27 |
| Component drug group 4 | 322.738±75.927 |
Hepatic tissue collagen staining fibrosis statistical result by stages sees Table 4.With normal rat relatively, 4 all rat model liver tissue fibrosis degree obviously raise (P＜0.01).Compare with 4 all model group, traditional decoction and component drug have tangible reduction effect to the liver tissues of rats fibrosis.Each component drug group liver tissues of rats collagen staining fibrosis slightly is better than the traditional decoction group by stages, but does not have significant difference between the two.
Table 4 is respectively organized the medicine group to DMN hepatic fibrosis rats hepatic fibrosis influence by stages.
Effect embodiment 2 Chinese medicine compositions of the present invention are analyzed the hepatic tissue pathology section of the treatment effect of N-nitrosodimethylamine (DMN) model
Microscope photographing is carried out in the hepatic tissue pathology section of respectively organizing among the effect embodiment 1 after medicine is treated the DMN hepatic fibrosis rats, see Fig. 2.
Wherein, No. 4 prescription groups are the component drug group 4 in effect embodiment 1 table 1, and full side group is the traditional decoction group.
Conclusion is as follows:
Shown that by above result of study component drug group and traditional decoction group all can be improved serological index, wherein component drug is better than the traditional decoction group.In addition, traditional decoction and component drug have tangible reduction effect to the liver tissues of rats fibrosis, and each component drug group rat hydroxyproline index and hepatic tissue collagen staining fibrosis all are better than the traditional decoction group by stages.
Claims (15)
1. a Chinese medicine composition is characterized in that comprising following ingredients: Herba Artemisiae Scopariae extract, Fructus Gardeniae extract, Radix Et Rhizoma Rhei extract I and Radix Et Rhizoma Rhei extract II; Wherein, the ratio of weight and number of described Herba Artemisiae Scopariae extract, Fructus Gardeniae extract, Radix Et Rhizoma Rhei extract I and Radix Et Rhizoma Rhei extract II is 1: 0.2: 0.2: 0.2～1: 10: 15: 15, above ratio was the ratio of weight and number of each extract dry weight;
In the described Herba Artemisiae Scopariae extract, the organic acid chemical compound accounts for the quality percentage composition of Herba Artemisiae Scopariae extract dry weight greater than 8%, and wherein, chlorogenic acid accounts for the quality percentage composition of Herba Artemisiae Scopariae extract dry weight more than or equal to 3%; In the described Fructus Gardeniae extract, jasminoidin accounts for the quality percentage composition of Fructus Gardeniae extract dry weight more than or equal to 40%; Among the described Radix Et Rhizoma Rhei extract I, rhubarb polyoses accounts for the quality percentage composition of Radix Et Rhizoma Rhei extract I dry weight greater than 40%; Among the described Radix Et Rhizoma Rhei extract II, rhatannin accounts for the quality percentage composition of Radix Et Rhizoma Rhei extract II dry weight more than or equal to 8%, and wherein, gallic acid accounts for the quality percentage composition of Radix Et Rhizoma Rhei extract II dry weight more than or equal to 1%.
2. Chinese medicine composition as claimed in claim 1, it is characterized in that: the ratio of weight and number of described Herba Artemisiae Scopariae extract, Fructus Gardeniae extract, Radix Et Rhizoma Rhei extract I and Radix Et Rhizoma Rhei extract II is 1: 0.2: 0.2: 0.2～1: 5: 5: 5, above ratio was the ratio of weight and number of each extract dry weight.
3. Chinese medicine composition as claimed in claim 2, it is characterized in that: the ratio of weight and number of described Herba Artemisiae Scopariae extract, Fructus Gardeniae extract, Radix Et Rhizoma Rhei extract I and Radix Et Rhizoma Rhei extract II is 1: 1: 1: 1～1: 3: 10: 10, above ratio was the ratio of weight and number of each extract dry weight.
4. as each described Chinese medicine composition of claim 1～3, it is characterized in that: in the described Herba Artemisiae Scopariae extract, the quality percentage composition that the organic acid chemical compound accounts for the Herba Artemisiae Scopariae extract dry weight is 10%～15%, wherein, to account for the quality percentage composition of Herba Artemisiae Scopariae extract dry weight be 4.7%～6% to chlorogenic acid; In the described Fructus Gardeniae extract, the quality percentage composition that jasminoidin accounts for the Fructus Gardeniae extract dry weight is 55%～60%; Among the described Radix Et Rhizoma Rhei extract I, the quality percentage composition that rhubarb polyoses accounts for Radix Et Rhizoma Rhei extract I dry weight is 62%～65%; Among the described Radix Et Rhizoma Rhei extract II, the quality percentage composition that rhatannin accounts for Radix Et Rhizoma Rhei extract II dry weight is 10%～12%, and wherein, the quality percentage composition that gallic acid accounts for Radix Et Rhizoma Rhei extract II dry weight is more than or equal to 2%.
5. as each described Chinese medicine composition of claim 1～4, it is characterized in that: described Herba Artemisiae Scopariae is dry aerial parts or its processed product of feverfew BINHAO or Herba Artemisiae Scopariae; Described Fructus Gardeniae is dry mature fruit or its processed product of Maguireothamnus speciosus Fructus Gardeniae; Described Radix Et Rhizoma Rhei is dry root, rhizome or its processed product of polygonum rheum palmatum, Rheum tanguticum or Rheum officinale.
6. as each described Chinese medicine composition of claim 1～5, it is characterized in that: the raw material of preparation Herba Artemisiae Scopariae extract, Fructus Gardeniae extract and two kinds of Radix Et Rhizoma Rhei extracts, promptly the ratio of weight and number of Herba Artemisiae Scopariae, Fructus Gardeniae and Radix Et Rhizoma Rhei is 1: (0.1-2): (0.1-2).
7. as each described Chinese medicine composition of claim 1～6, it is characterized in that: described Herba Artemisiae Scopariae extract is made by following method:
(1) be that 0%～30% ethanol water joins in the extraction equipment with capillary wormwood medical material and percent by volume, heating and refluxing extraction 0.5～2 hour, ethanol water is 10～25 times of capillary wormwood quality of medicinal material, extract 1～3 time, make the capillary wormwood extracting solution after filtration, be evaporated to 1～3 times of capillary wormwood medical material volume;
(2) add ethanol water in the capillary wormwood extracting solution that step (1) makes, the percent by volume of ethanol water is 60%～80%, places 10～14 hours, centrifugal, filtration obtains clear liquid, is evaporated to 1～3 times of capillary wormwood medical material volume, makes the capillary wormwood concentrated solution;
(3) the capillary wormwood concentrated solution that step (2) is made is pH to 2～5 of the hydrochloric acid solution adjusting capillary wormwood concentrated solution of 0.1～2mol/L with concentration, filters, and makes capillary wormwood filtrate;
(4) the capillary wormwood filtrate that step (3) is made, through absorption with macroporous adsorbent resin, earlier with the water elution impurity of 2～3 times of column volumes and discard cleanout fluid, 3～6 times of column volumes of reuse, percent by volume are 40～60% ethanol water eluting, behind the eluent concentrate drying, make Herba Artemisiae Scopariae extract.
8. as each described Chinese medicine composition of claim 1～7, it is characterized in that: described Fructus Gardeniae extract is made by following method:
(1 ') with the cape jasmine fruit coarse powder with the ethanol water of percent by volume 50%～80% heating and refluxing extraction 15～60 minutes in extraction equipment, ethanol water is 8～12 times of crude drug coarse powder quality, extract 1～3 time, obtain gardenia extract through centrifugal, filtration, be evaporated to 3～5 times of crude drug volume;
(2 ') will extract concentrated solution through absorption with macroporous adsorbent resin by the Fructus Gardeniae of step (1 ') gained, elder generation is 0～10% ethanol water eluting impurity with percent by volume, the back will promptly get Fructus Gardeniae extract with the ethanol water eluting of percent by volume 25%～35% behind the eluent concentrate drying.
9. Chinese medicine composition as claimed in claim 8 is characterized in that: used percent by volume is that the use amount of 0～10% ethanol water is 3～6 times of column volumes when carrying out eluting in the step (2 ').
10. as each described Chinese medicine composition of claim 1～9, it is characterized in that: described Radix Et Rhizoma Rhei extract I is made by following method:
(1 ") Radix Et Rhizoma Rhei water or ethanol water are carried out reflux, extract,, with extracting liquid filtering, get supernatant, promptly get Radix Et Rhizoma Rhei extract;
The Radix Et Rhizoma Rhei extract that (2 ") make step (1) is concentrated into volume mass with crude drug than being 0.5-3ml/g; add volumn concentration 95% ethanol water and reach volumn concentration 60%～80% to containing the alcohol amount; left standstill 8～24 hours; filter; with ethanol water washing precipitation; drying promptly gets Radix Et Rhizoma Rhei extract I.
11. Chinese medicine composition as claimed in claim 10 is characterized in that: step (1 ") in, the volumn concentration of described ethanol water is greater than 0, is less than or equal to 30%; The time of described reflux, extract, is 10-60 minute, and the number of times of extraction is 1-3 time.
12. Chinese medicine composition as claimed in claim 10 is characterized in that: step (2 ") in, the described temperature that leaves standstill is 2～10 ℃; The volumn concentration of the ethanol water of washing precipitation is 50%～95%.
13. as each described Chinese medicine composition of claim 1～12, it is characterized in that: described Radix Et Rhizoma Rhei extract II is made by following method: with the supernatant concentration that obtains after filtering in claim 10 or the 12 described steps (2 "), drying gets final product.
14. treat and/or prevent application in the medicine of hepatic disease in preparation as each described Chinese medicine composition of claim 1～13.
15. application as claimed in claim 14 is characterized in that: described hepatic disease is jaundice or the hepatic fibrosis that hepatitis, cholestasis cause.
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