CN104042675A - Compound agaricus blazei murill polysaccharide composition and application thereof - Google Patents
Compound agaricus blazei murill polysaccharide composition and application thereof Download PDFInfo
- Publication number
- CN104042675A CN104042675A CN201410317140.1A CN201410317140A CN104042675A CN 104042675 A CN104042675 A CN 104042675A CN 201410317140 A CN201410317140 A CN 201410317140A CN 104042675 A CN104042675 A CN 104042675A
- Authority
- CN
- China
- Prior art keywords
- agaricus blazei
- blazei murrill
- compound recipe
- polysaccharide
- radix
- 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.)
- Pending
Links
Abstract
The invention discloses a compound agaricus blazei murill polysaccharide composition and application thereof. The composition is prepared from the following traditional Chinese medicinal materials in parts by weight: 5-15 parts of agaricus blazei murill, 15-45 parts of astragalus membranaceus, 5-18 parts of scutellaria baicalensis, 10-38 parts of mushroom, 10-30 parts of isatis root, 10-20 parts of honeysuckle and 10-35 parts of liquorice. Testing results indicate that the compound agaricus blazei murill polysaccharide composition has a stronger effect on inhibiting salmonella, weaker on escherichia coli, and weakest on streptococcus suis, and the MIC (minimum inhibitory concentration) is respectively 0.075g/mL, 0.075g/mL and 0.3g/mL. Pharmacological experiments prove that the compound agaricus blazei murill polysaccharide oral solution does not have remarkable lesion on organs, is low in toxicity and large in safety range, and has the function of remarkably enhancing the body immunity. Therefore, the compound agaricus blazei murill polysaccharide composition has a large potential when being developed into clinical administration.
Description
Technical field
The invention belongs to Chinese medicine extraction technical field, particularly relate to compound recipe Agaricus Blazei Murrill polysaccharide compositions and application thereof.
Background technology
China's natural resources of Chinese medicinal materials is abundant, wherein finds that there is plurality of Chinese and has the effect that improves immunity of organisms.For example Radix Rhodiolae extract rhodioloside not only can promote the maturation of dendritic cell and the ability of its antigen presentation, and cell that can be inducing sustained and humoral immunization (Zhao etal, 2013); Herba Andrographis has the ability that improves Peritoneal Cells of Mice, neutrophilic granulocyte; Gansu Radix Codonopsis can strengthen by D-galactose, induced large age Mus immunologic function energy.By a large amount of clinical practices of research worker, show, the drug supporting healthy energy with the effect of raising immunity of organism has: Radix Codonopsis, the Radix Astragali, Radix Angelicae Sinensis etc.; The antipyretic and antidotal type medicine that strengthens reticuloendothelial system cytophagy ability has: Rhizoma Coptidis, Folium Isatidis, Herba Houttuyniae etc.; The Chinese medicine that can improve T cell ratio has: Radix Codonopsis, Radix Ginseng, the Rhizoma Atractylodis Macrocephalae etc.; The Chinese medicine that can promote body antibody to generate has: Flos Lonicerae, Semen Cuscutae, Radix Et Caulis Acanthopanacis Senticosi etc.
Along with the development of constantly bringing forth new ideas of Chinese medicine, traditional Chinese medical science scholar according to monarch, minister, help, make the principles of formulating prescriptions, work out the multiple vigorate qi and replenish the blood that has, improve the prescription of immunity of organisms.Chinese medical science research discovery, the Chinese medicine of many types has significant bacteriostasis.For example Fructus Tsaoko extract is 0.75mg/ml to staphylococcus aureus MIC, to colibacillary MIC, is 1.5mg/ml; Wide No. nine eucalyptus leaf essential oils of woods to the MIC of staphylococcus aureus, escherichia coli, bacillus subtilis be respectively 10,20,20ml/L; Herba Taraxaci Content of Chlorogenic Acid has stronger inhibitory action to staphylococcus aureus, escherichia coli, dysentery bacterium etc., its MIC is respectively 0.0250g/ml, 0.0063g/mL, 0.0125g/mL, black stone ear crude extract has certain inhibitory action (Wang Bin, 2013) to staphylococcus aureus, escherichia coli, bacillus subtilis.
That Chinese medicine has is pollution-free, do not produce the features such as fastbacteria, but its complicated component, the composition with bacteriostasis is less.The antipathogenic composition of different Chinese medicines is not quite similar, and general conclusion has the compositions such as flavone, alkaloid, quinones.Soybean isoflavone is better than gram negative bacteria to the inhibitory action of gram positive bacteria, to staphylococcus aureus MIC, is that the wild Actinidia arguta Sieb.et Zucc total flavones of 0.2mg/mL has inhibitory action to staphylococcus aureus, escherichia coli, rhizopus etc.
The present invention contains Agaricus blazei Murrill, the Radix Astragali, Lentinus Edodes, Radix Isatidis, Flos Lonicerae, Radix Scutellariae, Radix Glycyrrhizae by selected to the compound recipe Agaricus Blazei Murrill polysaccharide compositions of developing, their composition extract has been carried out to the inhibition test research of Salmonella, escherichia coli, Streptococcus suis, to providing foundation for clinical practice.
Summary of the invention
The invention discloses a kind of compound recipe Agaricus Blazei Murrill polysaccharide compositions, it is characterized in that it is comprised of the Chinese crude drug of following portions by weight:
Agaricus blazei Murrill 1-20 Radix Astragali 1-50
Radix Scutellariae 2-20 Lentinus Edodes 1-50
Radix Isatidis 5-30 Flos Lonicerae 2-25
Radix Glycyrrhizae 1-40.
Preferred Chinese crude drug of the present invention forms:
Agaricus blazei Murrill 1-15 Radix Astragali 1-45
Radix Scutellariae 2-18 Lentinus Edodes 1-38
Radix Isatidis 5-30 Flos Lonicerae 2-20
Radix Glycyrrhizae 1-35.
Another preferred Chinese crude drug of the present invention forms:
Agaricus blazei Murrill 5-15 Radix Astragali 15-45
Radix Scutellariae 5-18 Lentinus Edodes 10-38
Radix Isatidis 10-30 Flos Lonicerae 10-20
Radix Glycyrrhizae 10-35.
The present invention further discloses compound recipe Agaricus Blazei Murrill polysaccharide compositions, it is comprised of compositions wherein compound recipe Agaricus Blazei Murrill polysaccharide extract and pharmaceutically acceptable pharmaceutical carrier.Described compositions is that oral formulations is or injection.Oral formulations is wherein tablet, pill, oral liquid, capsule, powder or granule; Injection is aseptic parenteral solution or liquid spray, drop, injection, automatic injector assembly or suppository.
The present invention further discloses the preparation method of compound recipe Agaricus Blazei Murrill polysaccharide compositions, it is characterized in that being undertaken by following step:
(1) extraction of Agaricus Blazei Murrill polysaccharide
Take Agaricus blazei Murrill mushroom 10kg, grinding particle size is 40 orders, and after pulverizing, 500g keeps sample, and 9.5kg dry powder is used as extraction polysaccharide.Material-water ratio is 1:20, and 3.5kg Agaricus blazei Murrill dry powder is than 70kg water, and extraction temperature is 100 ℃, and extraction time is 3h; The centrifugal 10min of 2500r/min, concentrating under reduced pressure; 95% ethanol precipitate with ethanol concentrated solution, precipitate with ethanol is than being 1:3, the precipitate with ethanol time is 24h; Polysaccharide precipitation is dried.
(2) extraction of astragalus polysaccharides
Take Radix Astragali 50g, grinding particle size is 40 orders, and dry powder adds 8 times of water gagings to soak 12h, and reflux, extract, 2h, leaches solution, and medical material adds the water reflux, extract, 2h of 5 times of amounts again, discards filtering residue merging filtrate, the concentrated concentrated solution 50mL that obtains; Concentrated solution carries out ultracentrifugation, and the filtrate of separating and filtering is also carried out 75% ethanol precipitate with ethanol, the standing 12h of room temperature; Precipitation is dry.
(3) extraction of lentinan
Take Lentinus Edodes 50g, grinding particle size is 40 orders, and dried thin mushroom powder is pressed 1:25 solid-liquid ratio, in 95 ℃ of water-bath lixiviate 3h, extracts twice; Sucking filtration, supernatant adds 3 times of 95% ethanol, and alcohol is analysed; Centrifugal alcohol is analysed liquid, and gained is deposited in 50 ℃ of air dry ovens dry.
(4) compound recipe Agaricus Blazei Murrill polysaccharide extract:
By recipe quantity, Radix Scutellariae, Radix Isatidis, Radix Glycyrrhizae are decocted with water to 1.5h, filter, the another device of filtrate is collected; Flos Lonicerae and above-mentioned medicinal residues decoct with water secondary, each 0.5h, collecting decoction, standing filtration; Filtrate decompression is concentrated into about 800mL, and concentrated solution adds Agaricus Blazei Murrill coarse polysaccharide 1g, Lentinus Edodes crude polysaccharides 2g, and Radix Astragali crude polysaccharides 2g, had both obtained compound recipe Agaricus Blazei Murrill polysaccharide extract;
(5) the pharmaceutically acceptable pharmaceutical carrier of compound recipe Agaricus Blazei Murrill polysaccharide extract being mixed and made into oral formulations is or injection.
The Agaricus blazei Murrill that the present invention selects is to originate in the medicinal and edible fungus of the class of South America Brazil, and its mouthfeel is good, delicious flavour.The effects such as that Agaricus blazei Murrill water extract polysaccharide has is antibacterial, antitumor, antioxidation, blood sugar lowering, raising immunity of organisms.Agaricus Blazei Murrill polysaccharide be combined to Mice Body with fructose 1,6-diphosphate in lactic acid dehydrogenase have remarkable potentiation, can improve body resisting fatigue ability.
Astragalus polysaccharides is the water solublity heteropolysaccharide extracting from the medical herbs Radix Astragali, has the effects such as antitumor, antioxidation, raising immunity of organisms.Astragalus polysaccharides can increase specificity hepatitis B virus antibody and T cell-proliferation activity, meanwhile, can stimulate the maturation of dendritic cell, is the potential adjuvant of hepatitis B virus DNA vaccine.The solid tumor that astragalus polysaccharides can significantly suppress H22 hepatocarcinoma in BALB/c mouse body increases, spleen, thymus index and macrophage phagocytic ability also significantly improve, meanwhile, can promote the secretion of IL-2 in serum, IL-12, TNF-α, thereby suppress the deterioration of tumor.Astragalus polysaccharides suppresses CD4 in vivo
+c and D25
+t cell proliferation and its concentration and time is relation in gradient, and astragalus polysaccharides may be by recovering the expression of FOXp3 in cytokine balance and people's tumor microenvironment of hepatocellular carcinoma.
Lentinus Edodes is Basidiomycetes Lentinus Edodes Pseudomonas Lentinus Edodes fungus, is medicinal double edible fungi.On the books to Lentinus Edodes in ancient books: QI invigorating is not hungry, control wind removing blood stasis and stomach reinforcing helps food.Modern medicine proves, lentinan has remarkable efficacy improving immunity of organisms and anticancer aspect.Lentinan is combined the apoptosis that human mouth squamous cell cancerous cell can be significantly induced in use with S-1, reduce the expression of TS and DPDmRNA, and the expression of rising OPRTmRNA, has obvious cancer resistant effect.
Flos Lonicerae is the plant alabastrum part of Radix Ophiopogonis, and its sweet in the mouth is cold in nature, in the traditional Chinese medical science for heat-clearing and toxic substances removing, wind-dispelling heat-dissipating.Modern medicine is for inhibiting bacteria and diminishing inflammation, antiallergic, adjusting carbohydrate metabolism etc.Lonicera japonica and ethanol extract thereof have antiallergic characteristic.
Chinese medicine Radix Isatidis belongs to the root of cruciferae isatis, is recorded in the earliest in the > > of < < Sheng Nong's herbal classic, has heat-clearing and toxic substances removing, the effect of removing heat from blood sore-throat relieving.Modern medicine shows, active component sterols material, alkaloid and the polysaccharide of Radix Isatidis have the effects such as antiviral, antibacterial, antiinflammatory, antiendotoxin, antioxidation.
Radix Glycyrrhizae has another name called Herba Hedyotis cantonensis, is the rhizome of leguminous plant, light fruit and Glycyrrhiza inflata Bat..It is flat that it is recorded in the neutral sweet in the mouth of < < Sheng Nong's herbal classic > > the earliest, and GUIXIN, lung, spleen channel, have the effects such as heat-clearing and toxic substances removing, eliminating phlegm and stopping cough, invigorating the spleen and replenishing QI.Modern medicine study shows, the active component of Radix Glycyrrhizae is mainly flavonoid and saponins compound, has the effects such as antiviral, antiinflammatory, antibacterial, blood sugar lowering.
Radix Scutellariae, has another name called Radix Scutellariae, Labiatae Scutellaria, and its bitter in the mouth is cold in nature, has the effect of heat clearing and damp drying, hemostasis, eliminating fire and detoxication.Modern medicine shows, the effect such as that Radix Scutellariae has is anticancer, antibacterial, antioxidation, anti-hemolysis.Balcalin extract can suppress staphylococcus aureus growth, its MIC > 2048 μ g/mL, and can suppress the α HL hemolytic activity on staphylococcus aureus culture, be concentration dependence, but can not suppress the expression of α HL.It also has the mitigation to the A549 cell injury of staphylococcus aureus induction.
The curative effect situation of test is as follows:
| Test number | Salmonella | Escherichia coli | Streptococcus suis |
| Test 1 | 0.075g/mL | 0.075g/mL | 0.3g/mL |
Conclusion:
(1) forming the different consumption of identical selection has a certain impact to curative effect
(2) finally the formula of choosing consists of:
Agaricus blazei Murrill 1-15 Radix Astragali 1-45
Radix Scutellariae 2-18 Lentinus Edodes 1-38
Radix Isatidis 5-30 Flos Lonicerae 2-20
Radix Glycyrrhizae 1-35.
The present invention further discloses compound recipe Agaricus Blazei Murrill polysaccharide extract antibacterial, improve shown good effect aspect immunity of organisms.Disclosed compound recipe Agaricus Blazei Murrill polysaccharide compositions, it is comprised of compound recipe Agaricus Blazei Murrill polysaccharide extract and pharmaceutically acceptable pharmaceutical carrier.Can be made into oral formulations or injection.For example can be according to conventional medicine preparation technique by pharmaceutical composition, using polysaccharide powder as active component, closely mix to prepare pharmaceutical composition of the present invention with pharmaceutically suitable carrier, this carrier can for example, according to the extensive various dosage form of the form for administration of wanting (oral or parenteral as intramuscular) employing.When preparing the compositions of peroral dosage form, any common drug medium all can be used.Therefore for oral liquid as suspension, elixir and solution, suitable carrier and additive comprise water, ethylene glycol, oil, ethanol, flavoring agent, antiseptic, stabilizing agent, coloring agent etc.; For oral solid formulation, as powder, capsule, Caplet, soft capsule and tablet, suitable carrier and additive comprise starch, sucrose, diluent, granulating agent, lubricant, binding agent, disintegrating agent etc.Because be easy to administration, Tablet and Capsula agent represents the oral presented in unit dosage form of most convenient, obviously uses solid medicinal carrier in such situation.If needed, tablet can pass through standard technique sugar coating or enteric coating.For parenteral admistration, carrier comprises sterilized water conventionally, for example also can comprise in order to reach as hydrotropy object or for other compositions of anticorrosion use.Also suspension for injection can be prepared, in such situation, suitable liquid-carrier, suspending agent etc. can be used.
The every dosage unit of pharmaceutical composition herein for example sheet, capsule, powder, injection etc. should comprise the essential effective ingredient amount that discharges effective dose as above.The every dosage unit of pharmaceutical composition herein for example sheet, capsule, powder, injection, suppository etc. should comprise approximately 10～500 mg and can be with approximately 0.01～5.0 mg/kg/day, more preferably from about 0.1～0.5 mg/kg/day, more preferably from about 0.28g/Kg-2.8g/Kg.Yet, dosage can according to experimenter's needs, the severity of the disease for the treatment of and the compound of use change.Administration every day or all after date increments can be used.
Preferably these compositionss are in presented in unit dosage form, for oral, parenteral, intranasal, Sublingual or rectally or for sucking or spraying administration, this presented in unit dosage form is tablet, pill, oral liquid, capsule, powder, granule, aseptic parenteral solution or liquid spray, drop, injection, automatic injector assembly or suppository for example.Or, compositions can by main active ingredient and pharmaceutically suitable carrier for example common tabletting composition as corn starch, lactose, sucrose, sorbitol, Pulvis Talci, stearic acid, magnesium stearate, dicalcium phosphate or glue and other medicinal diluents for example water mix, to form the compositions that comprises the present invention's design.The novel present composition in liquid form can merge for oral or drug administration by injection, syrup, water or the oil suspension that this liquid form comprises aqueous solution, suitable flavouring with contain edible oil for example Emulsion and the elixir and similar pharmaceutical carrier of the flavouring of cottonseed oil, Oleum sesami, Oleum Cocois or Oleum Arachidis hypogaeae semen.Suitable dispersant or suspending agent for aqueous suspension comprise synthetic and natural glue, for example tragakanta, arabic gum, alginate, dextran, sodium carboxymethyl cellulose, methylcellulose, polyvidon or gelatin.For example, for the oral administration of tablet or capsule form, active drug composition can be with oral, nontoxic pharmaceutical acceptable inert carriers as ethanol, glycerol, water etc. combination.In addition,, when hope or while needing, suitable binding agent, lubricant, disintegrating agent and coloring agent also can be incorporated in mixture.Suitable binding agent comprises, is not limited to starch, gelatin, natural sugar if glucose or beta lactose, Semen Maydis sweetener, natural and synthetic glue are as arabic gum, tragakanta or enuatrol, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride etc.Disintegrating agent comprises, is not limited to starch, methylcellulose, agar, Bentonite, xanthan gum etc.
The daily dose of preferred composition changes at every per day for adults 0.01～1,000 mg of wide region.Oral administration, pharmaceutical composition is preferably with tablet form administration, the effective ingredient that it comprises 0.01,0.05,0.1,0.5,1.0,2.5,5.0,10.0,15.0,25.0,50.0,100,150,200,250 and 500 mg, for the dose titration of treated patient's symptom.Medicine effective quantity conventionally with every day approximately 0.001 mg/kg～approximately 5 mg/kg body weight dosage level supply with, preferred scope is that compound recipe Agaricus Blazei Murrill polysaccharide oral liquid contains crude drug 0.56g/mL (adult's dosage 16.8g/60kg body weight), and compositions also can be by the scheme administration of 1～4 time every day.Those skilled in the art can easily determine ideal occlusion pharmaceutical quantities, and these dosages change the progress of the concentration of the extract along with concrete use, mode of administration, preparation, mode of administration and disease disease.The factor relevant to treated concrete patient, comprises that patient age, body weight, diet and administration time can cause adjusting dosage in addition.
The present invention is more detailed to be described below:
First:
The processing technology of compound recipe Agaricus Blazei Murrill polysaccharide compositions (oral liquid is representative) and accelerated stability test research
1 material and instrument
1.1 test material
Agaricus blazei Murrill, the Radix Astragali, Lentinus Edodes, Radix Isatidis, Flos Lonicerae, Radix Scutellariae, Radix Glycyrrhizae
98% concentrated sulphuric acid (Tianjin Sheng Miao Fine Chemical Co., Ltd), anthrone reagent (Shanghai Shen Xiang chemical reagent company limited), glucose (AR) (National Institute for Food and Drugs Control).
1.2 test apparatus
Huge peak high speed disintegrator, Tianjin Chinese Medicine Machinery Plant manufactures; DZFZ9 fully automatic electric hot steam generator, Jiatian Manufacturing Co., Ltd., Shanghai; LD4-40 Beijing medical centrifuge; Concentrating under reduced pressure instrument, the safe light industry of fine horse; RE-52AA Rotary Evaporators, Shanghai Ya Rong biochemical equipment Instrument Ltd.; DHG-9143 air dry oven, Shanghai Fuma Experiment Equipment Co., Ltd..
Spectrophotometer (T6 new century); DHG-9143 air dry oven; Sartorius PB-10P
hmeter; XMT-DA constant water bath box.
2 test methods
The extraction of 2.1 polysaccharide
2.1.1 the extraction of Agaricus Blazei Murrill polysaccharide
Take 20 kilograms of Agaricus blazei Murrill mushrooms, grinding particle size is 40 orders, pulverizes latter 0.5 kilogram and keeps sample, and 9.5 kilograms of dry powder are used as extraction polysaccharide.Material-water ratio is 1:20, and 3.5 kilograms of Agaricus blazei Murrill dry powder are than 701 kg water, and extraction temperature is 100 ℃, and extraction time is 3h; Centrifugal 10 minutes of 2500r/min, concentrating under reduced pressure; 95% ethanol precipitate with ethanol concentrated solution, precipitate with ethanol is than being 1:3, the precipitate with ethanol time is 24h; Polysaccharide precipitation is dried.
2.1.2 the extraction of astragalus polysaccharides
Take Radix Astragali 50g, grinding particle size is 40 orders, and dry powder adds 8 times of water gagings to soak 12h, and reflux, extract, 2h, leaches solution, and medical material adds the water reflux, extract, 2h of 5 times of amounts again, discards filtering residue merging filtrate, the concentrated concentrated solution 50mL that obtains; Concentrated solution carries out ultracentrifugation, and the filtrate of separating and filtering is also carried out 75% ethanol precipitate with ethanol, the standing 12h of room temperature; Precipitation is dry.
2.1.3 the extraction of lentinan
Take Lentinus Edodes 50g, grinding particle size is 40 orders, and dried thin mushroom powder is pressed 1:25 solid-liquid ratio, in 95 ℃ of water-bath lixiviate 3h, extracts twice; Sucking filtration, supernatant adds 3 times of 95% ethanol, and alcohol is analysed; Centrifugal alcohol is analysed liquid, and gained is deposited in 50 ℃ of air dry ovens dry.
The processing technology of 2.2 compound recipe Agaricus Blazei Murrill polysaccharide oral liquids
Radix Scutellariae, Radix Isatidis, Radix Glycyrrhizae decoct with water 1.5h, filter, and the another device of filtrate is collected; Flos Lonicerae and above-mentioned medicinal residues decoct with water secondary, each 0.5h, collecting decoction, standing filtration; Filtrate decompression is concentrated into about 800mL, and concentrated solution adds Agaricus Blazei Murrill coarse polysaccharide 1g, Lentinus Edodes crude polysaccharides 2g, and Radix Astragali crude polysaccharides 2g, adds distillate to 1000ml, fill, autoclaving and get final product.
The preparation of 2.3 glucose standard solution
Precision takes 105 ℃ of dextrose standard sample 100mg that are dried to constant weight, with distilled water, dissolves and is settled to 100mL.Measure 20mL and be settled to 100mL, obtaining concentration is the glucose standard solution of 0.2mg/mL.
The preparation of 2.4 anthrone reagents
Precision takes anthrone 0.2g, adds 80% concentrated sulphuric acid 100mL, after dissolving completely, is placed in brown bottle matching while using.
The drafting of 2.5 standard curves
Measure successively dextrose standard sample solution 0.2,0.4,0.6,0.8,1.2mL is placed in 10mL dry test-tube, distinguish adding distil water to 2mL, the anthrone reagent that adds 4mL now to join along tube wall, 100 ℃ of water-bath 10min, be cooled to after room temperature, standing 20min, take anthrone reagent as blank, surveys absorbance in 620nm place, take absorbency Y as vertical coordinate, concentration X is abscissa, drawing standard curve (Guo Xiaolei etc., 2010).
The mensuration of 2.6 sugared content
With pipette, extract 2mL oral liquid in 10mL dry test-tube, the anthrone reagent that adds 4mL now to join along tube wall, 100 ℃ of water-bath 10min, be cooled to after room temperature, standing 20min, take anthrone reagent as blank, in 620nm place, survey absorbance (Zhang Hong etc., 2012).
2.7 accelerated test
Get 20130315,20130316, the oral liquid of 20,130,317 3 kind of different batches, simulation listing packing, be placed in 40 ℃ ± 2 ℃, under 75% ± 5%RH condition, place 6 months, duration of test the 1st, 2,3,6 the end of month sampling and testing compound recipe Agaricus Blazei Murrill polysaccharide oral liquid character, the uniformity, P
h, polyoses content.
3 result of the tests
The physical characteristic of 3.1 polysaccharide extract rates and compound recipe Agaricus Blazei Murrill polysaccharide oral liquid
Agaricus Blazei Murrill coarse polysaccharide extraction ratio is 14.95%; Radix Astragali coarse polysaccharide extractive rate is 6.16%; The extraction ratio of Lentinus Edodes crude polysaccharides is 10.66%; Gained compound recipe Agaricus Blazei Murrill polysaccharide oral liquid is brown liquid, solution homogeneous, has aromatic odor.
The drafting of 3.2 standard curves and the foundation of regression equation
The standard curve of drawing, by the linear regression analysis of data, obtains the regression equation of polysaccharide concentration-absorbance: y=0.0476x+0.0001, coefficient R
2=0.9971, illustrate that equation is linear good.
3.3 accelerated tests change pH value
In test, the compound recipe Agaricus Blazei Murrill polysaccharide oral liquid pH value of 0,1,2,3,6 month is measured, result is respectively 4.21 ± 0.25,4.25 ± 0.17,4.22 ± 0.21,4.20 ± 0.40,4.12 ± 0.67.Analysis of variance, the 6th month with 0 month relatively, pH value have significant difference (
p< 0.05).
3.4 accelerated tests change polyoses content
In test, the compound recipe Agaricus Blazei Murrill polysaccharide oral liquid polyoses content of 0,1,2,3,6 month is measured, result is respectively 15.12.00 ± 0.00,20.72 ± 9.70,20.72 ± 9.70,20.72 ± 9.70,34.72 ± 4.84.Analysis of variance, the 6th month with 0 month relatively, polyoses content have significant difference (
p> 0.05).
3.5 character and the uniformity change, and in the accelerated test of 6 months, compound recipe Agaricus Blazei Murrill polysaccharide oral liquid is brown liquid, uniformity homogeneous.
4 discuss
Stability test is the pith of study of tcm new drug, is to investigate medicament time dependent rule under the factors such as temperature, humidity, and wherein accelerated test is at 40 ℃ ± 2 ℃, under 75% ± 5%RH condition, evaluates one of important stability test of pharmacy quality.Accelerated test can within a short period of time, is the quality evaluation of new medicament, and storage requirement etc. provide test basis.
The Study on Acceleration of 6 months has been carried out in test to compound recipe Agaricus Blazei Murrill polysaccharide oral liquid, draw its character, and significant change was not all once all occurring, and 6th month pH value and polyoses content and 0 month are relatively, property that there were significant differences (
p< 0.05).Wherein 6th month pH value reduces, but do not surpass prescribed limit (permitted true etc., 1996); Polyoses content difference is not remarkable.
Experimental study shows, compound recipe Agaricus Blazei Murrill polysaccharide oral liquid accelerated to have good stability in storage test process at 6 months.
Second portion:
The non-clinical safety evaluation evaluation of compound recipe Agaricus Blazei Murrill polysaccharide compositions (oral liquid is representative)
1. rat clinical symptom examination
Observe medication group and control rats body weight change, clinical symptoms, death condition within 9 week experimental period; Observe every day, weighs weekly 1 time, and keep a record.
1.2 organ index inspections
After drug withdrawal, slaughter total Test rat, carry out careful system postmortem comprehensively, the organ that notes abnormalities answers emphasis to carry out histopathologic examination, core dirty, liver, spleen, lungs, kidney, brain, testis (male) and ovary (female) weighed, calculate organ coefficient (organ coefficient=Organ weight/the weight of animals * 100%) (Fei Chenzhong etc., 2013).
2.2.4 routine blood test and blood biochemical are learned and are checked
Before off-test, each treated animal fasting 12 h, freely drink water, and next day, eye socket was got hematometry hematological indices with etherization rat; Femoral artery blood sampling, with the centrifugal dirty separation of serum of 3500 r/min, measures blood parameters.
1.3 pathological examination
Get heart, liver, spleen, lungs, kidney, brain, thymus, ovary or the testis of respectively organizing rat, with 10% formalin solution, be fixed, paraffin embedding, HE dyeing, microscopy, takes a picture.
1.4 statistical analysis
Data with
± SD represents, adopts SPSS to analyze.
2 result of the tests
2..1 the acute toxicity tests
2.2 LD
50measurement result
In trial test feeding process, do not observe dead mouse and toxicity performance, execution Organs of Mice has no significantly observes pathological changes.Because being subject to the impact of drug level and gavage volume not measure LD
50, point out this oral liquid safety range very large, so done the maximum dosage-feeding test of mice.
2.3 maximum dosage-feeding result of the tests
Maximum dosage-feeding is: 67.2g/kg, 240 times of the dosage that is equivalent to be grown up.Whole mouse survivals in maximum dosage-feeding test, hair color gloss, activity freely, breathe normal, without ataxia, feed in order, without diarrhoea abdominal distention etc., administration group a few days ago produces obvious black feces in administration.
2.4 cut open inspection pathological change result
Cut open inspection mice, have no drug-induced obvious pathological change, observe normal.
The impact of 2.5 compound recipe Agaricus Blazei Murrill polysaccharide Oral Liquid On Mice body weight
Test as shown in Table 1, was weighed before on-test, and two groups of Mouse Weight differences are not remarkable.During off-test, two groups of Mouse Weight differences are not remarkable yet, and medicine is not obvious on Mouse Weight impact.
The impact of table 1 compound recipe Agaricus Blazei Murrill polysaccharide oral liquid Oral Liquid On Mice body weight
| Grouping | Quantity/only | First body weight/g | End-body weight/g |
| Administration group | 10 | 21.89±1.64 a | 31.57±4.02 a |
| Matched group | 10 | 21.29±1.73 a | 31.01±3.38 a |
Note: with column data shoulder mark lower case identical table differential different not remarkable (P>0.05), different expression significant differences (P<0.05) or extremely significantly (P<0.01).
The impact of 3.1 compound recipe Agaricus Blazei Murrill polysaccharide Oral Liquid On Mice Biochemical Indices In Serums
Result of the test as shown in Table 2, two groups of mice serum ALT, AST, Urea, there is not significant difference in Crea level.
The impact of table 2 compound recipe Agaricus Blazei Murrill polysaccharide oral liquid Oral Liquid On Mice Biochemical Indices In Serum
Note: with column data shoulder mark lower case identical table differential different not remarkable (P>0.05), different expression significant differences (P<0.05) or extremely significantly (P<0.01).
3.2 chronic toxicity test results
3.2.1 the clinical symptoms of rat
Duration of test, high, medium and low dosage group and control rats mental status, behavioral activity are good, and hair smoothing, without Folium Pini back of a bow diarrhoea phenomenon, is ingested and drinks water all normal.
3.2.2 the impact of compound recipe Agaricus Blazei Murrill polysaccharide oral liquid on rat body weight
Gavage 9 weeks, the rat body weight of the high, medium and low dosage group of compound recipe Agaricus Blazei Murrill polysaccharide oral liquid, with matched group comparison, without significant difference (table 3).
Each test group of table 3 and matched group body weight determination result (n=20)
Note: with column data shoulder mark lower case identical table differential different not remarkable (P>0.05), different expression significant differences (P<0.05) or extremely significantly (P<0.01).
3.2.3 the impact of compound recipe Agaricus Blazei Murrill polysaccharide oral liquid on Rats Organs and Tissues index
From organ index result, draw, in high dose group, organ index all increases, and its cardiac, liver, kidney and matched group comparison, have the significance difference opposite sex; In middle dosage group, organ index all increases, and its cardiac, liver and matched group comparison have the significance difference opposite sex (P < 0.05); In low dose group, lungs organ index reduces, and other all increase, with matched group comparison, all without the significance difference opposite sex (P > 0.05) (table 4).
Each test group of table 4 and matched group organ index measurement result (n=20)
Note: with column data shoulder mark lower case identical table differential different not remarkable (P>0.05), different expression significant differences (P<0.05) or extremely significantly (P<0.01).
3.2.4 the impact of compound recipe Agaricus Blazei Murrill polysaccharide oral liquid on rat serum routine
From blood routine examination result, can find out, in high dose group, each hemogram index raises, and wherein WBC, LYM, MID, GRA, PLT and matched group comparison, has the significance difference opposite sex (P < 0.05); In middle dosage group, RBC, HGB index reduce, and all the other all raise, and wherein WBC, GRA, PLT and matched group comparison has the significance difference opposite sex (P < 0.05); In low dose group, LYM index reduces, and all the other all raise, with matched group comparison, all without the significance difference opposite sex (P > 0.05) (table 5).
Each test group of table 5 and matched group routine blood test measurement result (n=20)
Note: with column data shoulder mark lower case identical table differential different not remarkable (P>0.05), different expression significant differences (P<0.05) or extremely significantly (P<0.01).
3.2.5 the impact of compound recipe Agaricus Blazei Murrill polysaccharide oral liquid on rat blood biochemical indicator
After administration 9 weeks, in high dose group, each biochemical indicator all raises, wherein Cre, TG and matched group comparison, have significant difference (
p< 0.05); In middle dosage group, AST index reduces, and all the other all raise, wherein TG index and matched group comparison, have the significance difference opposite sex (
p< 0.05); In low dose group, each biochemical indicator all raises, wherein TG index and matched group comparison, have the significance difference opposite sex (
p< 0.05) (table 6).
Each test group of table 6 and matched group Biochemical Indexes result (n=20)
Note: with column data shoulder mark lower case identical table differential different not remarkable (P>0.05), different expression significant differences (P<0.05) or extremely significantly (P<0.01).
3.2.6 pathological examination
Get heart, liver, spleen, lungs, kidney, brain, thymus, testis or the ovary of test group and all rats of matched group, observe internal organs without obvious pathological change.With 10% formalin solution, be fixed, carry out histopathologic examination, result shows: 3 dosage groups and matched group comparison, and without obviously pathological change.
4 discuss
4.1 chronic toxicity tests of compound recipe Agaricus Blazei Murrill polysaccharide oral liquid to rat
Chronic toxicity test is to be less than the chemicals of threshold dose to contact with laboratory animal for a long time, observe it to the whether toxigenous test of laboratory animal, being the core content of the non-clinical type safety evaluatio of medicine, is that medicine enters the important step of clinical trial from study of pharmacy.In test, medicine used is Chinese medicine compound, it follows " monarch " compatibility, the compound recipe consisting of compatibility is compared with single medicinal material, be improved curative effect, reduce toxic and side effects, take main symptom as main, take into account the mass action (Zhang Qi etc. of accompanied symptoms, 2007), there is the superiority such as antiinflammatory action in various degree.For example, the determining the pathogenesis of ZANG-FU disease based on the differentiation of symptoms and signs of Chinese medicine opinion is controlled has good therapeutic effect to chronic kidney disease, and has renal function protective effect.
pan etc. (2013) study and show, herbal mixture has materially affect for Cure of depression.Chinese medicine compound has good fungistatic effect.
Test is carried out the gavage of 60 days by a definite date and various test indexes is detected Wister rat, result shows, compound recipe Agaricus Blazei Murrill polysaccharide oral liquid on the mental status of rat, behavioral activity, hair luster, ingest and drink water without impact, illustrate that compound recipe Agaricus Blazei Murrill polysaccharide oral liquid is to the general metabolism of rat and physiological activity nonhazardous effect.
Body weight can directly reflect the nutriture of animal body and indirectly reflect the physiological situation of body, be an important indicator of chronic poison test.In test, the body weight of 3 dosage groups and matched group no significant difference, illustrate that compound recipe Agaricus Blazei Murrill polysaccharide oral liquid is to Growth in Rats unrestraint effect.
The organ index of animal is an important tests observation index of pharmacology and toxicologic study.The relative size of organ index, can illustrate the power of its function to a certain extent.Agaricus Blazei Murrill polysaccharide has a protective effect to liver is dirty; Astragalus polysaccharides can make liver cell arrange closely in order, and between Kupffer's cells core lobule, arteriovenous increases, and contributes to strengthen liver function; Astragalus polysaccharides can improve IL-1 β content in Cyprinus carpio kidney, and Promote immunity is replied.In test, heart, liver exponent increase in the heart in high dose group, liver, kidney and middle dosage group, with matched group relatively there were significant differences property, illustrate that compound recipe Agaricus Blazei Murrill polysaccharide oral liquid has facilitation to enhancing organ function.
It is to distribute by observing number change and form that routine blood test detects, judgement disease.In test, WBC, GRA, the PLT of the WBC in high dose group, LYM, MID, GLA, PLT and middle dosage group increase, with matched group relatively there were significant differences property, but all in range of normal value or approach normal value, this may be because Agaricus Blazei Murrill polysaccharide has improved immunity of organism, stimulates the propagation of lymphocyte, mononuclear cell etc.
To be respectively xenobiotic transform and the main place of metabolism at body for liver and kidney.The enzymes activity such as plasma A LT, AST can reflect liver cell damage situations and serious journey.Creatinine and blood urea nitrogen are the important indicators of reflection kidney bead filtering function damage.In test, AST, the ALT of 3 dosage groups all raise, but compare no significant difference with matched group, illustrate that liver is not subject to obvious damage.In pathological examination, 3 dosage groups and matched group comparison, have no notable difference, illustrate compound recipe Agaricus Blazei Murrill polysaccharide oral liquid to internal organs without obvious pathological changes.In sum, compound recipe Agaricus Blazei Murrill polysaccharide oral liquid toxicity is low, and safety range is large, has potentiality to be exploited.
Third part: compound recipe Agaricus Blazei Murrill polysaccharide compositions (take typical oral liquid as representative) In Vitro Bacteriostasis research
1 material and instrument
1.1 test material
Compound recipe Agaricus Blazei Murrill polysaccharide compositions, is prepared by TanJin Agricultural College veterinary test chamber.Main component has Agaricus Blazei Murrill polysaccharide, astragalus polysaccharides and Flos Lonicerae.
Salmonella, escherichia coli, Streptococcus suis (CVCC606 2 types) provides (can externally provide) by TanJin Agricultural College Preventive Veterinary Medicine laboratory
Tryptone (lot number: HB8270), yeast powder (lot number: HB8273), sodium chloride (lot number: HB8112), agar powder (lot number: HB8220) purchase by Qingdao GaoKeYuan Hai Bo Bioisystech Co., Ltd;
Nutrient agar (lot number: HG3492-93), be purchased from Tianjin Bei Lian fine chemicals development corporation, Ltd..
1.2 test apparatus
DHG-9143 air dry oven, Shanghai Fuma Experiment Equipment Co., Ltd.; SKY-110WX constant-temperature shaking culture device; SW-CJ-ID superclean bench; BCD-215KAZ refrigerator; JA2003 electronic balance, Shanghai Sunny Hengping Scientific Instrument Co., Ltd..
2 test methods
2.1 actication of culture
Sterile working is by Salmonella, escherichia coli, Streptococcus suis (CVCC606 2 types), and streak inoculation, in sterilized culture medium, is cultivated 24 h for 37 ℃ respectively.
The preparation of 2.2 bacteria suspensions
With inoculating loop, choose the bacterium colony that deactivates in LB fluid medium, cultivate 24 h for 37 ℃.
2.3 plate count
Get 8 sterilizing test tubes, number 1～8, add respectively the LB fluid medium of 3.6 mL sterilizings.Get 0.4 mL bacteria suspension stock solution and be added in the 1st test tube, mix, draw 0.4 mL and be added in the 2nd test tube, the rest may be inferred, and the 8th test tube discards 0.4 mL.Select 2 suitable dilution factors, get respectively 200 μ L and coat in LB solid medium, each dilution factor is made two plates, cultivates 24 h for 37 ℃.Obtain after each dull and stereotyped total plate count, obtain with dilution each dull and stereotyped average clump count, be multiplied by extension rate, calculate stock solution bacterial concentration (bacteria suspension concentration is generally 1.28 * 108 cfu/mL).
2.4 Oxford agar diffusion method drug sensitive tests
Get 200 μ L bacteria suspensions and be evenly coated on LB solid medium, the Oxford cup of sterilizing is positioned in culture medium, each culture medium is put three.By 6 mg/L, 3 mg/L, 1.5 mg/L, 0.75 mg/L, 0.375 mg/L, 0.1875 mg/L, 0.09375 mg/L, the oral liquid diluent of 0.046875 mg/L and blank liquid (distilled water) are added to respectively in the cup of Oxford, and each adds 150 μ L, and each concentration is done three repetitions.Be put in the constant incubator of 37 ℃ and cultivate, observed result after 24 h, and with slide gauge, inhibition zone is measured.
2.5 MIC methods
Medicinal liquid is done to 2 times of dilutions.Get 11 test tubes and add respectively 2 mL LB fluid mediums, get test medicinal liquid 2 mL and put into the 1st test tube, mix, then draw 2 mL mixing materials and put into the 2nd test tube, be diluted to successively the 9th test tube, from the 9th test tube, discard 2 mL mixing materials, the 10th test tube dosing do not add bacterium liquid, as negative control, the 11st test tube do not add medicinal liquid, as positive control.The bacteria suspension of preparation is diluted to 105 cfu/mL, is added in 10 test tubes, every test tube adds the bacterium liquid of 100 μ L, is placed in 37 ℃ of calorstats and cultivates 24 h.The culture of drawing respectively 200 μ L from 10 test tubes is evenly coated on LB solid medium, is placed in 37 ℃ of calorstats and cultivates 24 h, and the flat board of selecting bacterium colony to look into calculates its minimal inhibitory concentration (MIC).
2.6 criterion
Inhibition zone >=20 mm Gao Min; Quick in 10-19 mm; < 10 mm drug resistances.Minimum inhibitory concentration (MIC) refers to that medicine suppresses the least concentration of certain growth of microorganism completely, and it is basis for estimation that the clump count of take on plate is minimum inhibitory concentration (MIC) at the individual dilution drug level of minimum of 10-20.
3 result of the tests
3.1 drug sensitive test
From recording antibacterial circle diameter, can find out, compound recipe Agaricus Blazei Murrill polysaccharide compositions has certain inhibitory action to Salmonella, escherichia coli, Streptococcus suis, and wherein stronger to Salmonella inhibitory action, escherichia coli take second place, Streptococcus suis the most weak (table 7).
The inhibition of table 7 compound recipe Agaricus Blazei Murrill polysaccharide compositions to 3 kinds of antibacterials
3.2 MIC result of the tests
From result, compound recipe Agaricus Blazei Murrill polysaccharide compositions is 0.075 g/mL to Salmonella, colibacillary minimal inhibitory concentration, to the minimal inhibitory concentration of Streptococcus suis, is 0.3 g/mL.
The minimal inhibitory concentration of table 8 compound recipe Agaricus Blazei Murrill polysaccharide compositions to 3 kinds of antibacterials
Note: " ++ " represents that clump count is in 10; "+" represents that clump count is at 10-20; "-" represents that clump count is more than 20.
4 discuss
Compound recipe Agaricus Blazei Murrill polysaccharide compositions is to be formed by compatibilities such as Flos Lonicerae, Radix Scutellariae, Agaricus Blazei Murrill polysaccharide, astragalus polysaccharidess, and it is to bring into play drug effect by the mutual synergism between single medicinal material.Wherein, Flos Lonicerae, the Radix Astragali etc. is antipyretic and antidotal type Chinese medicine, and modern study shows, that such Chinese medicine has is antibacterial, the effect of opposing pathogenic microorganism and Promote immunity function.Polysaccharide is that the degree of polymerization surpasses 10 polysaccharide, has important pharmacological action; Astragalus polysaccharides has certain inhibitory action to escherichia coli, Salmonella and bacillus pyocyaneus; Particularly bacillus subtilis, staphylococcus aureus and escherichia coli are had to good inhibition.In test, compound recipe Agaricus Blazei Murrill polysaccharide compositions has shown inhibition in various degree to escherichia coli, Salmonella and Streptococcus suis.Wherein, compound recipe Agaricus Blazei Murrill polysaccharide compositions is better than Streptococcus suis to the inhibitory action of escherichia coli and Salmonella.
Conclusion:
Compound recipe Agaricus Blazei Murrill polysaccharide compositions is that to take Agaricus Blazei Murrill polysaccharide, astragalus polysaccharides, Flos Lonicerae etc. be main component, take and improve the new Chinese medicine compound that body's immunity is object, this research has been made thoroughly evaluating to compound recipe Agaricus Blazei Murrill polysaccharide composition production processing technology, accelerated stability, the safety evaluation of non-clinical property, raising immunity of organisms and In Vitro Bacteriostasis effect, draws to draw a conclusion:
(1), in compound recipe Agaricus Blazei Murrill polysaccharide composition production processing technology, show that the extraction ratio of Agaricus blazei Murrill, the Radix Astragali, Lentinus Edodes crude polysaccharides is respectively 14.95%, 6.16%, 10.66%.
(2) in compound recipe Agaricus Blazei Murrill polysaccharide composition stable Journal of Sex Research, show, after the accelerated test of 6 months, compound recipe Agaricus Blazei Murrill polysaccharide composition stable is good.
(3) in compound recipe Agaricus Blazei Murrill polysaccharide compositions, show in to acute toxicity test in mice and rat chronic toxicity test, compound recipe Agaricus Blazei Murrill polysaccharide compositions does not have toxicity to body, and safety range is large, has potentiality to be exploited.
(4) in compound recipe Agaricus Blazei Murrill polysaccharide compositions, improve in immunity of organisms test and show, compound recipe Agaricus Blazei Murrill polysaccharide compositions all has facilitation to the humoral immunization of body and non-characteristic immunity.
(5) outside compound recipe Agaricus Blazei Murrill polysaccharide composition, in fungistatic effect research, show, compound recipe Agaricus Blazei Murrill polysaccharide compositions is stronger to Salmonella inhibition, escherichia coli take second place, and Streptococcus suis is the most weak, and its MIC is respectively 0.075g/mL, 0.075g/mL, 0.3g/mL.
The good effect that compound recipe Agaricus Blazei Murrill polysaccharide compositions disclosed by the invention has is:
(1) compound recipe Agaricus Blazei Murrill polysaccharide composition production technique is simple, easily be used, test is studied by stability, safety and pharmacology pharmacodynamic that compound recipe Agaricus Blazei Murrill polysaccharide compositions is carried out, shown the significant medical value of new compound Chinese medicine, and new compound Chinese medicine is cheap, be 22.61 yuan/L as calculated, market potential is huge.
The specific embodiment
Below in conjunction with embodiment, the present invention is described, the scheme of embodiment described here, do not limit the present invention, one of skill in the art can make improvements and change according to spirit of the present invention, these described improvement and change all should be considered as in scope of the present invention, and the requirement of all having the right of scope of the present invention and essence limits; Wherein said Chinese medicinal herbs, reagent used are by commercially available.
Embodiment 1
The preparation of compound recipe Agaricus Blazei Murrill polysaccharide extract
(the extraction of 10 Agaricus Blazei Murrill polysaccharides
Take 10 kilograms of Agaricus blazei Murrill mushrooms, grinding particle size is 40 orders, pulverizes latter 0.5 kilogram and keeps sample, and 9.5 kilograms of dry powder are used as extraction polysaccharide.Material-water ratio is 1:20, and 7 jin of Agaricus blazei Murrill dry powder are than 701 kg water, and extraction temperature is 100 ℃, and extraction time is 3h; Centrifugal 10 minutes of 2500r/min, concentrating under reduced pressure; 95% ethanol precipitate with ethanol concentrated solution, precipitate with ethanol is than being 1:3, the precipitate with ethanol time is 24 hours; Polysaccharide precipitation is dried.
(2) extraction of astragalus polysaccharides
Take Radix Astragali 50g, grinding particle size is 40 orders, and dry powder adds 8 times of water gagings to soak 12h, and reflux, extract, 2h, leaches solution, and medical material adds the water reflux, extract, 2h of 5 times of amounts again, discards filtering residue merging filtrate, the concentrated concentrated solution 50mL that obtains; Concentrated solution carries out ultracentrifugation, and the filtrate of separating and filtering is also carried out 75% ethanol precipitate with ethanol, the standing 12h of room temperature; Precipitation is dry.
(3) extraction of lentinan
Take Lentinus Edodes 50g, grinding particle size is 40 orders, and dried thin mushroom powder is pressed 1:25 solid-liquid ratio, in 95 ℃ of water-bath lixiviate 3h, extracts twice; Sucking filtration, supernatant adds 3 times of 95% ethanol, and alcohol is analysed; Centrifugal alcohol is analysed liquid, and gained is deposited in 50 ℃ of air dry ovens dry.
(4) processing technology of compound recipe Agaricus Blazei Murrill polysaccharide
| Formula | Radix Scutellariae | Radix Isatidis | Flos Lonicerae | Radix Glycyrrhizae |
| （1） | 10 | 15 | 8 | 20 |
| （2） | 8 | 20 | 10 | 15 |
| （3） | 15 | 8 | 15 | 8 |
| （4） | 10 | 10 | 20 | 10 |
| （5） | 4 | 15 | 10 | 15 |
The formula medical material of above-mentioned (1)-(5) is decocted with water to secondary, each 0.5h, collecting decoction, standing filtration; Filtrate decompression is concentrated into about 800mL, and concentrated solution adds Agaricus Blazei Murrill coarse polysaccharide 1g, Lentinus Edodes crude polysaccharides 2g, and Radix Astragali crude polysaccharides 2g, mix homogeneously, had both obtained compound recipe Agaricus Blazei Murrill polysaccharide extract;
(5) the pharmaceutically acceptable pharmaceutical carrier of compound recipe Agaricus Blazei Murrill polysaccharide extract being mixed and made into oral formulations is or injection.
Embodiment 2:
Get compound recipe Agaricus Blazei Murrill polysaccharide powder 10g, glucose 50g water, complements to 1000ml.Preparation method: get the compound recipe Agaricus Blazei Murrill polysaccharide powder of recipe quantity, room temperature adds part water for injection, after stirring evenly, adds glucose, stirring and dissolving, then mend water for injection to 1000ml, and filter, embedding, sterilizing, obtains 5ml specification compound recipe Agaricus Blazei Murrill polysaccharide injection.
Embodiment 3:
Get compound recipe Agaricus Blazei Murrill polysaccharide powder 10g, mannitol 100g, sodium chloride 90g, water for injection complements to 1000ml.Preparation method: the compound recipe Agaricus Blazei Murrill polysaccharide powder of getting recipe quantity, under room temperature, be dissolved in appropriate water for injection, after stirring and dissolving, add mannitol, stirring and dissolving, mend again water for injection to 1000ml, through No. 6 incipient fusion filter degerming, filter, by filtrate fill under aseptic condition immediately, every 5ml, after lyophilization, seal, obtain compound recipe Agaricus Blazei Murrill polysaccharide lyophilized powder.
Embodiment 4:
Get compound recipe Agaricus Blazei Murrill polysaccharide extract 15g.Medical starch 30.0g, magnesium stearate 2.5g, 50% appropriate amount of ethanol, granulates, and granulate is dried, dress 2# capsule, every containing compound recipe Agaricus Blazei Murrill polysaccharide 0.1g.
Embodiment 5:
Get compound recipe Agaricus Blazei Murrill polysaccharide powder 50g, medical starch 30.0g, dextrin 30.0g, 50% appropriate amount of ethanol, is fully uniformly mixed granulation by above-mentioned raw materials, at 60～70 ℃, is dried 2～4 hours, film-making, every containing compound recipe Agaricus Blazei Murrill polysaccharide 0.5g.
Embodiment 6
Get compound recipe Agaricus Blazei Murrill polysaccharide 20g, the sterile deionized water dilution that adds 0.32L, (being that 20g polysaccharide powder is made about 320ml), cold preservation 24 hours, the centrifugal precipitation of going, final clear liquid adds xylitol 5g, and cold preservation is filtered, and obtains clear and bright filtrate, in clear liquid, add again sterile deionized water to 400ml, pack into and in aseptic bottle, make oral liquid, every 10 ml of loading amount.
After the preferred embodiment describing in detail, being familiar with this technology personage can be well understood to, can carry out various variations and modification not departing under above-mentioned claim and spirit, any simple modification, equivalent variations and modification that all foundations technical spirit of the present invention is done above embodiment, all belong to the scope of technical solution of the present invention.And the present invention is not also limited to the embodiment of example in description.
Claims (8)
1. a compound recipe Agaricus Blazei Murrill polysaccharide compositions, is characterized in that it is comprised of the Chinese crude drug of following portions by weight:
Agaricus blazei Murrill 1-20 Radix Astragali 1-50
Radix Scutellariae 2-20 Lentinus Edodes 1-50
Radix Isatidis 5-30 Flos Lonicerae 2-25
Radix Glycyrrhizae 1-40.
2. compound recipe Agaricus Blazei Murrill polysaccharide compositions claimed in claim 1, is characterized in that it is comprised of the Chinese crude drug of following portions by weight:
Agaricus blazei Murrill 1-15 Radix Astragali 1-45
Radix Scutellariae 2-18 Lentinus Edodes 1-38
Radix Isatidis 5-30 Flos Lonicerae 2-20
Radix Glycyrrhizae 1-35.
3. compound recipe Agaricus Blazei Murrill polysaccharide compositions claimed in claim 1, is characterized in that it is comprised of the Chinese crude drug of following portions by weight:
Agaricus blazei Murrill 5-15 Radix Astragali 15-45
Radix Scutellariae 5-18 Lentinus Edodes 10-38
Radix Isatidis 10-30 Flos Lonicerae 10-20
Radix Glycyrrhizae 10-35.
4. compound recipe Agaricus Blazei Murrill polysaccharide compositions described in claim 1-3 any one, it is comprised of compositions wherein compound recipe Agaricus Blazei Murrill polysaccharide extract and pharmaceutically acceptable pharmaceutical carrier.
5. compound recipe Agaricus Blazei Murrill polysaccharide compositions described in claim 4, compositions is wherein that oral formulations is or injection.
6. compound recipe Agaricus Blazei Murrill polysaccharide compositions described in claim 5, oral formulations is wherein tablet, pill, oral liquid, capsule, powder or granule; Injection is aseptic parenteral solution or liquid spray, drop, injection, automatic injector assembly or suppository.
7. the preparation method of compound recipe Agaricus Blazei Murrill polysaccharide compositions described in claim 1-3 any one, is characterized in that being undertaken by following step:
(1) extraction of Agaricus Blazei Murrill polysaccharide
Take Agaricus blazei Murrill mushroom 10kg, grinding particle size is 40 orders, and after pulverizing, 500g keeps sample, and 9.5kg dry powder is used as extraction polysaccharide, and material-water ratio is 1:20, and 3.5kg Agaricus blazei Murrill dry powder is than 70kg water, and extraction temperature is 100 ℃, and extraction time is 3h; The centrifugal 10min of 2500r/min, concentrating under reduced pressure; 95% ethanol precipitate with ethanol concentrated solution, precipitate with ethanol is than being 1:3, the precipitate with ethanol time is 24h; Polysaccharide precipitation is dried;
(2) extraction of astragalus polysaccharides
Take Radix Astragali 50g, grinding particle size is 40 orders, and dry powder adds 8 times of water gagings to soak 12h, and reflux, extract, 2h, leaches solution, and medical material adds the water reflux, extract, 2h of 5 times of amounts again, discards filtering residue merging filtrate, the concentrated concentrated solution 50mL that obtains; Concentrated solution carries out ultracentrifugation, and the filtrate of separating and filtering is also carried out 75% ethanol precipitate with ethanol, the standing 12h of room temperature; Precipitation is dry;
(3) extraction of lentinan
Take Lentinus Edodes 50g, grinding particle size is 40 orders, and dried thin mushroom powder is pressed 1:25 solid-liquid ratio, in 95 ℃ of water-bath lixiviate 3h, extracts twice; Sucking filtration, supernatant adds 3 times of 95% ethanol, and alcohol is analysed; Centrifugal alcohol is analysed liquid, and gained is deposited in 50 ℃ of air dry ovens dry;
(4) compound recipe Agaricus Blazei Murrill polysaccharide extract:
By recipe quantity, Radix Scutellariae, Radix Isatidis, Radix Glycyrrhizae are decocted with water to 1.5h, filter, the another device of filtrate is collected; Flos Lonicerae and above-mentioned medicinal residues decoct with water secondary, each 0.5h, collecting decoction, standing filtration; Filtrate decompression is concentrated into about 800mL, and concentrated solution adds Agaricus Blazei Murrill coarse polysaccharide 1g, Lentinus Edodes crude polysaccharides 2g, and Radix Astragali crude polysaccharides 2g, had both obtained compound recipe Agaricus Blazei Murrill polysaccharide extract;
(5) the pharmaceutically acceptable pharmaceutical carrier of compound recipe Agaricus Blazei Murrill polysaccharide extract being mixed and made into oral formulations is or injection.
8. the application of compound recipe Agaricus Blazei Murrill polysaccharide compositions in preparing anti-bacteria raising immunity of organisms medicine described in claim 1-3 any one.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410317140.1A CN104042675A (en) | 2014-07-07 | 2014-07-07 | Compound agaricus blazei murill polysaccharide composition and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410317140.1A CN104042675A (en) | 2014-07-07 | 2014-07-07 | Compound agaricus blazei murill polysaccharide composition and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104042675A true CN104042675A (en) | 2014-09-17 |
Family
ID=51496398
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410317140.1A Pending CN104042675A (en) | 2014-07-07 | 2014-07-07 | Compound agaricus blazei murill polysaccharide composition and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104042675A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104435130A (en) * | 2014-12-12 | 2015-03-25 | 广西科技大学 | Application of essential oil from leaves of eucalyptus urophylla E.grandis, Guanglin 9 as bacteriostatic drug |
| CN109922666A (en) * | 2016-08-26 | 2019-06-21 | 纽崔克荷兰有限责任公司 | The method of animal feed and this feed of offer |
| CN115364138A (en) * | 2022-09-19 | 2022-11-22 | 博斯创想(上海)医疗科技有限公司 | Compound bacterial liquid for improving chemotherapy side effects and preparation method thereof |
-
2014
- 2014-07-07 CN CN201410317140.1A patent/CN104042675A/en active Pending
Non-Patent Citations (6)
| Title |
|---|
| 彭勇胜等: "响应面法优化姬松茸多糖的提取工艺", 《现代食品科技》 * |
| 戴肖冬等: "姬松茸抑菌作用的初步研究", 《中国食用菌》 * |
| 赵翠莉等: "姬松茸抗病作用的研究进展", 《黑龙江畜牧兽医》 * |
| 郜鹏云等: "复方姬松茸多糖口服液对大鼠的慢性毒性试验", 《中国畜牧兽医》 * |
| 郜鹏云等: "姬松茸多糖口服液对小鼠的急性毒性研究", 《天津农学院学报》 * |
| 黄东杰等: "黄芪多糖的提取方法综述", 《企业科技与发展》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104435130A (en) * | 2014-12-12 | 2015-03-25 | 广西科技大学 | Application of essential oil from leaves of eucalyptus urophylla E.grandis, Guanglin 9 as bacteriostatic drug |
| CN109922666A (en) * | 2016-08-26 | 2019-06-21 | 纽崔克荷兰有限责任公司 | The method of animal feed and this feed of offer |
| CN115364138A (en) * | 2022-09-19 | 2022-11-22 | 博斯创想(上海)医疗科技有限公司 | Compound bacterial liquid for improving chemotherapy side effects and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106942724A (en) | Composition acted on regulating intestinal canal Bacterial community and preparation method and application | |
| CN108991529B (en) | Food composition for invigorating spleen and nourishing vitality and preparation method thereof | |
| CN103800661A (en) | Composition for improving immunity and relieving physical fatigue and preparation thereof | |
| CN1326536C (en) | Pharmaceutical composition for treating and preventing respiratory tract viral infection, its preparation process and application | |
| CN106924477B (en) | Composite traditional Chinese medicine fermentation preparation produced by mixed bacteria fermentation and preparation method thereof | |
| CN104042675A (en) | Compound agaricus blazei murill polysaccharide composition and application thereof | |
| Li | Chinese herbal medicine | |
| CN114344351A (en) | Selenium-rich ganoderma lucidum spore powder capsule and preparation method thereof | |
| CN102058810B (en) | Lung-clearing asthma-relieving phlegm-reducing Chinese patent drug | |
| CN104288217B (en) | A kind of Chinese medicine composition and purposes | |
| CN105394515A (en) | Pleurotus sajor-caju-cattail solid-state fermented functional drink and preparation method | |
| CN104367673B (en) | A kind of Chinese medicine composition treating Eimeria species | |
| CN107893035A (en) | A kind of culture medium of Hericium erinaceus, sealwort bioconversion mycelium, the mycelial extract of sealwort bioconversion and application thereof | |
| CN108553504A (en) | Join stilbene powder and preparation method thereof | |
| CN107648330A (en) | A kind of hypoglycemic composition based on fennelflower and hypoglycemic anaesthetic | |
| CN104069177A (en) | Compound agaricus blazei polysaccharide oral liquid as well as preparation method and application thereof | |
| CN101062374B (en) | Method for preparing Chinese traditional combination capsule for treating cancer and the product thereof | |
| CN103417653B (en) | Veterinary-use compound pure Chinese herbal medicine for treating coccidium and preparation method of oral liquid of same | |
| CN105454956A (en) | A hericium erinaceus-common cattail solid fermentation functional drink and a preparing method | |
| CN102423384B (en) | Traditional Chinese drug preparation for treating lung cancer, and preparation method thereof | |
| CN101366852B (en) | Chinese medicinal herb for treating riemerella anatipestifer disease and preparation method thereof | |
| CN105454957A (en) | A pleurotus citrinopileatus-common cattail solid fermentation functional drink and a preparing method | |
| CN103191268B (en) | Traditional Chinese medicinal composition for treating lung cancer | |
| CN115715783B (en) | Traditional Chinese medicine composition for preventing and treating white diarrhea of pigs and preparation method thereof | |
| CN104013650B (en) | Bacillus licheniformis and herbal polysaccharide composition and its preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140917 |
|
| WD01 | Invention patent application deemed withdrawn after publication |