CN1778824A - Multiple homogeneous tremella polysaccharide, its extraction and medicinal composition with this compound as active components - Google Patents
Multiple homogeneous tremella polysaccharide, its extraction and medicinal composition with this compound as active components Download PDFInfo
- Publication number
- CN1778824A CN1778824A CN 200410072865 CN200410072865A CN1778824A CN 1778824 A CN1778824 A CN 1778824A CN 200410072865 CN200410072865 CN 200410072865 CN 200410072865 A CN200410072865 A CN 200410072865A CN 1778824 A CN1778824 A CN 1778824A
- Authority
- CN
- China
- Prior art keywords
- atf
- polysaccharide
- homogeneous
- tremella
- tremella polysaccharide
- 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
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Several homogeneous tremella polysaccharides, its extraction and medicinal composition as the compound as active ingredient are disclosed. The process is carried out by boiling tremella spore fermentative, depositing by alcohol, gradient extracting by different concentration NaOH solution, column chromatography separating, purifying to obtain base-extracted tremella polysaccharide ATF-A, ATF-B, ATF-C and water-extracted tremella polysaccharide WTF-A and WTF-B, and mixing homogeneous tremella polysaccharide with medicinal accessory material to obtain various preparations. It can decrease immunity, delay senility and have antineoplastic and radiation-resistant functions.
Description
Technical field
The present invention relates to several herbal polysaccharide materials, more particularly, is through extracting, separate the multiple homogeneous tremella polysaccharide that obtains from the Chinese medicine white fungus.Simultaneously also disclose its extracting method and be the pharmaceutical composition of activeconstituents, and said composition is in the application that is used for preparing medicines such as enhancing immunity, antitumor, anti-radiation, leukocyte increasing, antiviral, anti-ageing, AIDS resisting with this compound.
Background technology
White fungus (Tremella fuciformis Berk is abbreviated as TF) claims Tremella again, is a kind of higher fungi, belongs to the Heterobasidiomycetae Tremellaceae.White fungus is universally acknowledged precious edible mushrooms and important medicinal material, has many-sided biological activity, comprises that antitumor, antiviral, anti-inflammatory, radioprotective, immunomodulatory, promotion protein, biological nucleic acid are synthetic etc.
At present, existing Tremella fuciformis spores polysaccharide capsule and various white fungus healthcare products on the market, these are made by the white fungus crude extract, active constituent content is low, and impurity is many, and is water-soluble bad, particularly lower in the stomach and intestine specific absorption, the capsule of making, tablet oral result are relatively poor, do not have better therapeutic effect.In addition,, therefore still can not make injection, influence the normal performance of curative effect because tremella polysaccharide is only accomplished crude extract at present.On existing patent and the various magazine STUDY ON POLYSACHAROSE is confined on the water-soluble component mostly, and fewer for the research of slightly water-soluble component, the alkali of having delivered that has carries that skunk bush polysaccharide, alkali are carried the Marasmius equicrinis Muell. polysaccharide, alkali is carried indocalamus leaf polyose etc.
The present invention's water-soluble portion from tremella spore saleratus extracts and has obtained two kinds of equal one polysaccharide (WTF-A, WTF-B); Water-insoluble part from the tremella spore fermented product obtains 4 kinds of equal one polysaccharide (ATF-A, ATF-B, ATF-C, ATF-D) with the alkali lye extraction, is mixed polysaccharide, and particularly alkali lye extracts the tremella spore fermented product, has not yet to see any bibliographical information.
Summary of the invention
In view of the foregoing, the inventor has carried out a large amount of experimental studies to water-soluble component, the slightly water-soluble component of tremella spore fermented product, determined that water is carried, serial of methods such as alkali lye gradient extraction, separation, purifying tremella polysaccharide and structural analysis, thereby finished the invention of multiple homogeneous tremella polysaccharide.
First purpose of the present invention is, multiple homogeneous tremella polysaccharide is provided, and is called for short ATF-A, ATF-B, ATF-C, ATF-D, WTF-A, WTF-B.
Another object of the present invention is, the extracting method of multiple homogeneous tremella polysaccharide is provided.
A further object of the present invention is that providing with the multiple homogeneous tremella polysaccharide is the pharmaceutical composition of activeconstituents.
Also purpose of the present invention is, providing is that the pharmaceutical composition of activeconstituents is in the application that is used for preparing medicines such as enhancing immunity, antitumor, anti-radiation, leukocyte increasing, antiviral, anti-ageing, AIDS resisting with the multiple homogeneous tremella polysaccharide.
Multiple homogeneous tremella polysaccharide of the present invention, it be by the tremella spore fermented product through poach, alcohol precipitation, remove albumen or NaOH aqueous solution gradient extraction with different concns after, obtain content at the multiple homogeneous tremella polysaccharide more than 90% through column chromatography for separation, purifying; Mainly contain following composition: alkali desilver fungus polysaccharides ATF-A, ATF-B, ATF-C, ATF-D, water desilver fungus polysaccharides WTF-A, WTF-B; Analyze through the efficient gel column chromatography, their weight-average molecular weight is respectively 62830,63710,58962,51454,73000 and 68000; Wherein,
ATF-A mainly is made up of D-glucose, D-wood sugar, D-seminose;
ATF-B mainly is made up of D-glucose, D-wood sugar, D-seminose;
ATF-C mainly is made up of D-glucose, L-rhamnosyl;
ATF-D mainly is made up of D-glucose, D-wood sugar, D-seminose, D-semi-lactosi;
WTF-A mainly is made up of L-Fucose, D-wood sugar, D-seminose, D-semi-lactosi and D-glucose;
WTF-B mainly is made up of L-Fucose, D-wood sugar, D-seminose, D-glucose.
The extracting method of described multiple homogeneous tremella polysaccharide comprises the steps:
A. the extraction of water-soluble homogeneous tremella polysaccharide:
(1) tremella spore saleratus adds poach and carries, and the cooling back is centrifugal, adds ethanol sedimentation after the supernatant concentration.Cross the leaching precipitation, vacuum-drying gets Crude polysaccharides.
(2) Crude polysaccharides adds poach and carries, and the cooling back is centrifugal, and supernatant liquor removes albumen with the sevage method, removes the solution ethanol sedimentation behind the albumen.Cross the leaching precipitation, vacuum-drying must be made with extra care polysaccharide.
(3) get refining polysaccharide and separate through gel filtration chromatography, use the sodium chloride aqueous solution wash-out, elutriant concentrates and adds ethanol sedimentation, and vacuum-drying makes water and carries homogeneous tremella polysaccharide WTF-A, WTF-B.
B. the extraction of slightly water-soluble homogeneous tremella polysaccharide:
After going out water-soluble component with hot water extraction, remaining slightly water-soluble component is adopted the sodium hydroxide lye gradient extraction of different concns, after peracid neutralization, ethanol sedimentation, usefulness Sevage method are removed deproteinize, the cellulose column level separates and purifying, obtain four polysaccharide components, ATF-A, ATF-B, ATF-C, ATF-D are mixed polysaccharide. and through being accredited as equal one polysaccharide of the present invention, concrete operations are as follows:
(1) after going out water-soluble component with hot water extraction, with 0.1mol/L-10mol/LNaOH aqueous solution gradient extraction, hydrochloric acid is neutralized to neutrality to remaining slightly water-soluble component, be concentrated into small volume, with the flowing water dialysis, 70-90% ethanol precipitating makes alkali and carries Crude polysaccharides;
(2) alkali is carried Crude polysaccharides and is removed albumen through the sevage method, and with the flowing water dialysis, 70-90% ethanol precipitating must be made with extra care polysaccharide;
(3) will make with extra care cellulose chromatography separation on the polysaccharide, and with different concentrations of sodium chloride aqueous solution wash-out, collect different elutriants, and concentrate the back alcohol precipitation, vacuum-drying makes alkali and carries homogeneous tremella polysaccharide, ATF-A, ATF-B, ATF-C, ATF-D.
Described gel chromatography column comprises DEAE-Sephadex gel A-25 and Sephadex G-200, Sephadex G-100, Sephadex G-150.Described cellulose chromatography comprises DEAE-52-cellulose, DEAE-32-cellulose.
Described sevage method is removed albumen and is meant chloroform: the method for propyl carbinol=4: 1.
Of the present invention is the pharmaceutical composition of activeconstituents with the homogeneous tremella polysaccharide, is meant that said composition contains the homogeneous tremella polysaccharide for the treatment of significant quantity, and one or more pharmaceutically acceptable carrier.
Wherein, the treatment significant quantity of described homogeneous tremella polysaccharide is generally 0.1～500mg; Be preferably 1～200mg; Best 5～100mg.
Described composition comprises oral preparations, liquid preparation or external preparation.Wherein, oral preparations is tablet, capsule, slow releasing tablet, dripping pill or oral liquid; Liquid preparation is injecting and administering preparations such as freeze-dried powder, injection liquid, large and small transfusion or intravenous drip liquid; External preparation is creme or paste.
Described large and small transfusion is meant homogeneous tremella polysaccharide sodium-chlor or glucose injection agent.
Described pharmaceutically acceptable carrier, comprise thinner, weighting agent (as N.F,USP MANNITOL, lactose, polyoxyethylene glycol) conventional in the preparation, tackiness agent (starch, Microcrystalline Cellulose), disintegrating agent (as carboxymethyl cellulose, the low hydroxypropylcellulose that replaces), lubricant (as talcum powder, Magnesium Stearate), wetting agent (as propylene glycol, ethanol), oxidation inhibitor (as Sodium Pyrosulfite, Sulfothiorine), sanitas (phenylcarbinol); Stablizer (EDTA-2Na, Sulfothiorine, Sodium Pyrosulfite, S-WAT, thanomin, sodium bicarbonate, niacinamide) or the like.Above-mentioned auxiliary material can be a common dose, mixes with homogeneous tremella polysaccharide with proportioning commonly used, and after the homogeneous tremella polysaccharide consumption was determined, the proportioning between each pharmaceutical excipient can suitably be regulated as required.
The preparation concrete operations of various pharmaceutical preparations are as follows:
(1), takes by weighing and contain the 10-70% homogeneous tremella polysaccharide by the configuration total amount, the weighting agent of adding 30-90%, tackiness agent, disintegrating agent, lubricant, correctives etc., fully mix and make particle, compacting in 60-70 ℃ of dry 2-4 hour in flakes, make every to contain homogeneous tremella polysaccharide 40-240mg; Maybe with the wet granular made directly at 60-70 ℃ of dry 2-4 hour, be filled into then in the hungry area shell, make every capsules contain homogeneous tremella polysaccharide 30-280mg.
(2), the preparation of injection liquid and large and small transfusion: be the homogeneous tremella polysaccharide and the distilled water dosing of getting 0.01-70%, or add stablizer that after the G4 funnel filtered, the packing sterilization made.
Described freeze-dried be to mix by 0.08-50% homogeneous tremella polysaccharide and pharmaceutical excipient (combinations of one or more of lactose, N.F,USP MANNITOL, glycine, sorbyl alcohol, glucose), pharmaceutical excipient is a N.F,USP MANNITOL preferably, the packing sterilization makes.
(3), take by weighing and contain the 10%-60% homogeneous tremella polysaccharide, add the weighting agent, disintegrating agent, correctives of 40%-90% etc., fully mix with the 12-14 mesh sieve and make particle by the configuration total amount.At 60-70 ℃ of dry 2-4 hour, make every gram granule contain homogeneous tremella polysaccharide 100-300mg.
(4), take by weighing and contain the 10-60% homogeneous tremella polysaccharide by the configuration total amount, the weighting agent of adding 40%-90%, oxidation inhibitor etc., heating and melting temperature (40-90 ℃) makes raw material and auxiliary material fully miscible, drip and make ball, make every dripping pill contain homogeneous tremella polysaccharide 4-30mg.
Formulation rate of the present invention can be according to all multifactor adjustment such as route of administration, patient age, body weight, disease type and severity, and per daily dose is for being generally 0.1～300mg/kg; Be preferably 0.1～100mg/kg; Best .0.1～60mg/kg.Can be according to the clinical suitable change of pathology Signs.
Homogeneous tremella polysaccharide of the present invention (ATF-A, ATF-B, ATF-C, ATF-D, WTF-A, WTF-B) has following physicochemical property:
One, spectroscopic analysis:
1. infrared spectra;
Six kinds of pure product of polysaccharide are carried out fourier transform infrared spectrometry to be measured.The result shows, 3400cm
-1Broad peak be the stretching vibration of O-H and N-H, 2930cm
-1Be the absorption peak of C-H, these two groups of charateristic avsorption bands that the peak is a carbohydrate; Other 6 polysaccharide are at 1022cm
-1And 1150cm
-1All there is absorption the vicinity, illustrates that they all are pyranoses; Sample is at 850cm
-1All there is absorption the vicinity, illustrates to exist α type glycosidic link (being the α anomer) in the polysaccharide structures, and six polysaccharide are all α-pyranose.
2. UV scanning:
Shimadzu UV-3000 ultraviolet-visible spectrophotometer (day island proper Tianjin company).Six kinds of polysaccharide are water-soluble, be made into the solution of 10mg/50ml, carry out UV scanning, wavelength is 200nm-800nm.The scanning result sample does not all have to absorb at 260nm and 280nm place, illustrates in these six polysaccharide and does not all contain nucleic acid and protein.
Two, purity testing:
Gel column chromatography: get six kinds of each 15g of polysaccharide and be dissolved among the 0.9%Nacl of minimum volume, add Sephadex G-200 (2.5 * 33cm) capitals.Use the 0.9%Nacl wash-out, flow velocity 1.5ml/ hour, 1ml/ managed fraction collection, and the phenol sulfuric acid process is measured and respectively managed polysaccharide content, with 752 spectrophotometric determination absorbancys.The reaction that always is eluted to polysaccharide is negative, merge same composition, draw the relation curve of wash-out pipe number and absorbancy, the result shows, alkali desilver fungus polysaccharides ATF-A, ATF-B, ATF-C, ATF-D, the Sephadex G-200 gel filtration chromatography elution curve of water desilver fungus polysaccharides WTF-A, WTF-B is single symmetrical peak, illustrates that this sample is equal one polysaccharide.
Three, mean molecule flow measurement:
Liquid phase chromatogram condition:
Chromatographic column is TSK G 4000PWXL (6 * 300mm); Detector is Shodex; Column temperature is 35 ℃; Sample size is 20 μ l; Moving phase is 0.7%Na
2SO
4, flow velocity is 0.5ml/min.
The molecular weight of six kinds of polysaccharide is respectively ATF-A, ATF-B, ATF-C, ATF-D, their weight-average molecular weight of WTF-A, WTF-B is respectively 62830,63710,58962,51454,73000 and 68000.
Four, equal one polysaccharide composition measuring:
1, water-soluble equal one polysaccharide ply of paper is analysed:
(1) chromatographic paper: Whattman No1 chromatography filter paper.
(2) developping agent: propyl carbinol-Glacial acetic acid-water (4: 1: 5); Vinyl acetic monomer-pyridine-water (10: 4: 3)
(3) developer: aniline-phthalic acid (0.93g aniline, 1.66g phthalic acid are dissolved in the water saturated propyl carbinol of 100ml).
(4) method: each 8mg of sample thief WTF-(A-B), use 2N H respectively
2SO
4The 1mL tube sealing, 100 ℃ of hydrolysis 6hr, barium carbonate is neutralized to neutrality.Suction filtration is got filtrate, concentrates point sample.Launch for developping agent with propyl carbinol-Glacial acetic acid-water (4: 1: 5) and vinyl acetic monomer-pyridine-water (10: 4: 3), the corresponding standard product are done contrast, after launching to finish, volatilize developping agent, the spray developer.
2, the equal one polysaccharide of slightly water-soluble ply of paper is analysed
(1) chromatographic paper: Whatman No.1 chromatography filter paper
(2) developping agent: ethyl acetate-pyridine-water-Glacial acetic acid (5: 5: 3: 1)
(3) developer: aniline-phthalic acid (0.93g aniline and 1.66g phthalic acid are dissolved in the water saturated propyl carbinol of 100ml)
(4) method: get the pure product 10mg of each polysaccharide of ATF-(A-D) respectively and be dissolved in the 2m12mol/L trifluoroacetic acid, tube sealing, hydrolysis is 6 hours in the boiling water bath.Cooling back point sample,, expansion back spray developer saturated with above-mentioned developping agent done contrast with corresponding monose standard substance.
3, gas chromatographic analysis:
A. hydrolyzate is added 10mg oxammonium hydrochloride and 1ml anhydrous pyridine, warm dissolving is placed in 90 ℃ of water-baths heats 30min, be chilled to room temperature after, add 1ml anhydrous acetic acid acid anhydride, continue in this water-bath, to heat 30min, acetylize is finished, sample directly detects.And with this method with the acetylize of corresponding monose standard substance after sample presentation in the lump.
B. GC conditions:
Chromatographic column is a quartz capillary column, and stationary liquid is SE-52,30 meters of column lengths; Column temperature 180 ℃ keep 5min after per minute 3 degree that raise, keep 10min at 250 ℃; Detector is FID; The Sample Room temperature is 270 ℃; Carrier gas is N
2, flow velocity is 50ml/h.
C. gas chromatograph results:
ATF-A mainly is made up of D-glucose, D-wood sugar, D-seminose.
ATF-B mainly is made up of D-glucose, D-wood sugar, D-seminose.
ATF-C mainly is made up of D-glucose, L-rhamnosyl.
ATF-D mainly is made up of D-glucose, D-wood sugar, D-seminose, D-semi-lactosi.
WTF-A mainly is made up of L-Fucose, D-wood sugar, D-seminose, D-semi-lactosi and D-glucose.
WTF-B mainly is made up of L-Fucose, D-wood sugar, D-seminose, D-glucose.
The invention also discloses homogeneous tremella polysaccharide in the application that is used for preparing enhancing immunity, antitumor, anti-radiation, leukocyte increasing, antiviral, the anti-ageing medicine of waiting for a long time.
Following pharmacological evaluation has confirmed that homogeneous tremella polysaccharide has enhancing immunity, antitumor, anti-radiation, leukocyte increasing, pharmacological action such as antiviral.
One, antitumor action: tremella polysaccharide suppresses the lymphoma experiment
1 material
1.1 animal is selected:
The breeding IRM-2 of Institute of Radiation Medicine, Chinese Academy of Medical Sciences mouse inbred lines, the male and female dual-purpose.
1.2 knurl kind:
Animal housing of Institute of Radiation Medicine, Chinese Academy of Medical Sciences is from IRM-2 mouse inbred lines spontaneous lymphoma (pathology conclusion) cultivation of going down to posterity.
1.3 medicine preparation:
Get polysaccharide WTF-B, being mixed with concentration is 6mg/kg, 12mg/kg, and solvent is a distilled water.
2 methods:
2.1 inoculation:
Select tumor growth vigorous and do not have a diabrosis, the IRM-2 mouse that health condition is good, dislocation is put to death, be fixed on the operation plate, use the tincture of iodine, bromogeramine sterilization animal skin is shelled tumour in super clean bench under the aseptic condition, the incision tumour becomes the fritter about diameter 2mm, in the inguinal region place's subcutaneous vaccination of animal left side.
Therapeutic evaluation: put to death animal in 24 hours after the drug withdrawal, weigh, dissect and peel off the knurl piece, claim knurl heavy.
Tumor control rate: C-T/C * 100%
T is that the average knurl of administration group is heavy in the formula, and C is that the average knurl of control group is heavy
Experimental result:
| Group | Body weight (g) X ± (SD) | Heavy (g) X of knurl ± (SD) | Tumour inhibiting rate (%) | The P value | |||
| Beginning | Finally | Beginning | Finally | ||||
| ♂♀/2 | |||||||
| Contrast | 10 | 10 | 30.2±2.5 | 34.3±4.4 | 1.24±0.66 | ||
| 6mg/kg | 20 | 20 | 29.8±2.7 | 33.8±5.4 | 1.23±0.57 | 0.81％ | |
| 12mg/kg | 20 | 20 | 30.5±3.4 | 32.8±1.6 | 0.48±0.49 | 61.3％ | P＜0.01 |
The result shows that tremella polysaccharide is 61.3% to lymphadenomatous inhibiting rate when 12mg/kg.
Two. suppress lymphoma radiation synergism:
1 material
1.1 animal is selected:
The self-reproduction IRM-2 of Institute of Radiation Medicine, Chinese Academy of Medical Sciences mouse inbred lines, the male and female dual-purpose.
1.2 knurl kind:
Animal housing of Institute of Radiation Medicine, Chinese Academy of Medical Sciences is from IRM-2 mouse inbred lines spontaneous lymphoma (pathology conclusion) cultivation of going down to posterity.
1.3 medicine preparation:
Get polysaccharide WTF-A, being mixed with concentration is 6mg/kg, and solvent is a distilled water.
2 methods:
2.1 inoculation:
Select tumor growth vigorous and do not have a diabrosis, the IRM-2 mouse that health condition is good, dislocation is put to death, be fixed on the operation plate, use the tincture of iodine, bromogeramine sterilization animal skin is shelled tumour in super clean bench under the aseptic condition, the incision tumour becomes the fritter about diameter 2mm, in the inguinal region place's subcutaneous vaccination of animal left side.
2.2 grouping administration
Control group and r ray groups, medicine and r ray combination group are established with the animal random packet next day in the inoculation back, and be administered once every day, abdominal injection, and each 0.5ml, control group gives distilled water 0.5ml, successive administration 10 days.The r ray groups, every day is with the r radiation exposure of 1Gy 1 minute, Continuous irradiation 8 days.
2.3 therapeutic evaluation: put to death animal in 24 hours after the drug withdrawal, weigh, dissect and peel off the knurl piece, claim knurl heavy.
Tumor control rate: C-T/C * 100%
T is that the average knurl of administration group is heavy in the formula, and C is that the average knurl of control group is heavy
Polysaccharide (6mg/kg) and 1Gy r ray share and suppress the lymphoma experiment
| Group | Number of animals (only) | Body weight (g) X ± (SD) | Heavy (g) X ± (the S D) of knurl | Tumour inhibiting rate % | The P value | ||
| Beginning | Finally | Beginning | Finally | ||||
| Contrast | 10 | 10 | 30.2±2.5 | 33.4±4.4 | 1.24±0.66 | ||
| The r ray | 10 | 10 | 30.3±3.8 | 31.9±4.6 | 0.7±0.25 | 43.5 | P＜0.05 |
| Medicine+r ray | 10 | 10 | 30.5±3.8 | 31.5±2.8 | 0.34±0.32 | 72.6 | P＜0.001 |
The result shows, polysaccharide when concentration 6mg/kg and the r radiation exposure of 1Gy share, be 72.6% (P＜0.001) to the lymphoma inhibiting rate of IRM-2 mouse, compare significant difference with control group.
Three. radiation resistance:
1. experiment material
1.1 animal: purebred C57 mouse, body weight 22-24g.
1.2 experimental technique:
1.2.1 grouping and administration: animal is divided into four groups (15/group, male and female half and half) at random: control group, medicine (ATF-B) 6mg/kg, three dosage groups of 12mg/kg, 24mg/kg.Pre-irradiation continuous 3 administrations in the 3rd, 2,1 days, abdominal injection, 0.5ml/, control group is given and physiological saline, and abdominal injection, 0.5ml/ are only.
1.1.2 the condition of causing injury:
137Cs-gamma-rays one subtotal body irradiation, dosage is respectively: 7.005Gy and 7.5Gy, dose rate be 89.1404 human relations/minute.
1.1.3 observation index: (1) femur karyocyte is counted BMC (The number of nucleated cellsin bone marrow), and irradiation back was taken off vertebra with the mouse neck on the 9th day and put to death, and got a side femur, normal saline flushing marrow.Electron microscope is the numeration cell count down.
(2) the spleen tubercle is counted CFU (Colong forming unit of spleen) and spleen index, takes out spleen, weighs, and calculates spleen index.Spleen is put into BouinShi liquid, after 24 hours, the tubercle number (CFU-S) on the outstanding spleen surface of naked eyes numeration.
Experimental result:
Mouse is according to back the 9th day femur karyocyte number, spleen tubercle number and index and spleen index
| Group | BMC×10 6Femur/root | Heavy (the mg)/body weight (g) of spleen index spleen | CFU-S/spleen | Irradiation dose Gy |
| Control group 6mg/kg 12mg/kg 24mg/kg | 1.31±0.18 1.53±0.33 * 2.01±0.38 *** 1.73±0.49 ** | 0.993±0.206 1.115±0.196 * 1.084±0.188 1.134±0.186 | 1.29±1.14 2.80±3.2 *** 1.07±0.96 1.38±1.02 | 7.005 |
| Control group 6mg/kg 12mg/kg 24mg/kg | 1.795±0.28 2.27±0.44 ** 3.26±0.52 *** 3.34±0.75 *** | 1.03999±0.3341 1.1353±0.145 1.062±0.16 1.174±0.335 | 1.64±1.49 0.93±0.70 1.08±0.92 2.2±1.6 | 7.5 |
Annotate: * P＜0.05 * * P＜0.01 * * * P＜0.001
Tremella polysaccharide BMC when 6mg/kg, 12mg/kg, three dosage of 24mg/kg learns processing by statistics and has compared significant difference with control group.
Hemocytopoietic organ is the responsive organ of ray, and being subjected to the damage according to the back hemopoietic tissue is its most important pathological change, and the recovery of hemopoietic function is the key factor of prognosis.CFU-S system represents the polydirectional hemopoietic stem cell group of a class, the ability that they have self and break up to the red system of marrow, grain system and megalokaryocyte, and CFU-S is the main component that promotes that hematopoiesis recovers.BMC is the karyocyte composition in the marrow.The ability that on behalf of the body hemopoietic tissue, the variation of CFU-S and BMC recover.As can be seen, tremella polysaccharide can be protected the hemopoietic function of irradiation mouse, has radiation resistance from this experiment.
Four. leukogenic effect:
1. experiment material:
1.1 animal: purebred C57 mouse, body weight 22-24g.
1.2 experimental technique:
1.2.1 grouping and administration: animal is divided into five groups (15/group, male and female half and half) at random: control group, medicine (ATF-D) 6mg/kg, 15mg/kg, 30mg/kg, four dosage groups of 60mg/kg.Pre-irradiation continuous 3 administrations in the 3rd, 2,1 days, abdominal injection, 0.5ml/, control group is given and physiological saline, and abdominal injection, 0.5ml/ are only.
1.2.2 the condition of causing injury:
137Cs-gamma-rays one subtotal body irradiation, dosage is respectively: 7.5Gy, dose rate be 89.1404 human relations/minute.
Observation index: (1) leukocyte count: irradiation back was taken off vertebra with the mouse neck on the 9th day and is put to death, and eye socket is got blood, the white corpuscle diluted, and electron microscope is the numeration cell count down.
(2) dna content, irradiation back took off vertebra with the mouse neck on the 9th day puts to death, and gets a side femur, normal saline flushing marrow.Add reaction solution, measure absorbancy on the ultraviolet spectrophotometer.
(3) CFU-S (Colong forming unit of spleen) puts into BouinShi liquid with spleen, after 24 hours, and the tubercle number (CFU-S) on the outstanding spleen surface of naked eyes numeration
Experimental result:
Tremella polysaccharide is tested radiation injury mouse leukocyte increasing
| Group | DNA (OD/ root) | WBC×10 9/L | CFU-S |
| Contrast 6mg/Kg 15mg/Kg 30mg/Kg 60mg/Kg | 0.111±0.026 0.115±0.024 0.098±0.021 0.119±0.021 0.166±0.08 | 0.185±0.085 0.258±0.146 0.267±0.133 0.308±0.086 0.475±0.157 | 0.1±0.316 0.33±0.516 0.33±0.52 0.167±0.4 0.83±0.754 |
Learn by statistics and handle, 60mg/Kg group, DNA (OD/ root), WBC * 10
9/ L, the every index of CFU-S,
Compare significant difference with control group.
Five, strengthening immunity experiment
1. experiment material:
1.1 animal: purebred C57 mouse, body weight 22-24g.
1.2 experimental technique:
1.2.1 grouping and administration: animal is divided into five groups (15/group, male and female half and half) at random: control group, medicine 6mg/kg, 15mg/kg, four dosage groups of 30mg/kg, 60mg/kg.Pre-irradiation continuous 3 administrations in the 3rd, 2,1 days, abdominal injection, 0.5ml/, control group is given and physiological saline, and abdominal injection, 0.5ml/ are only.
1.2.3 the condition of causing injury:
137Cs-gamma-rays one subtotal body irradiation, dosage is respectively: 7.5Gy, dose rate be 89.1404 human relations/minute.
Observation index: (1) spleen index: take out spleen, weigh, calculate spleen index
(2) thymus index: take out thymus gland, weigh, calculate spleen index
Tremella polysaccharide improves the immunizing power experiment to the radiation injury mouse
| Group contrast 6mg/Kg 15mg/Kg 30mg/Kg 60mg/Kg | Spleen index 0.77 ± 0.376 0.89 ± 0.265 0.98 ± 0.244 0.91 ± 0.186 1.05 ± 0.06 | Thymus index 1.13 ± 0.294 1.07 ± 0.309 1.27 ± 0.246 1.29 ± 0.202 1.72 ± 0.375 |
Learn by statistics and handle, the 60mg/Kg group, spleen index, two indexs of thymus index have been compared significant difference with control group.
Six, antivirus action:
1. material
1.1 cell and virus
Tire ox pneumonocyte (Fetal Bovine Lung, FBL), bovine immunodeficiency virus R29 strain (BovineImmunodeficiency Virus R29 strain, BIVR29)
1.2 reagent and medicine
DMEM substratum, penicillin/streptomycin (P/S), Trypsin, Tris, EDTA, foetal calf serum (U.S. GIBCO/BRL company).AZT is available from Sigma company.
1.3 medicine
(1) positive control medicine AZT is dissolved in the dimethyl sulfoxide (DMSO) (DMSO), concentration 0.1mM (0.026mg/ml), and filtration sterilization is stored in-20 ℃.
(2) tremella polysaccharide (WTF-B) is diluted to three concentration of 0.1mg/ml, 0.15mg/ml 0.2mg/ml with PBS, and filtration sterilization is stored in-20 ℃.
2. method
2.1 cell cultures
With DEME substratum (1%P/S 10% foetal calf serum) at 37 ℃, 5%CO
2Cultivate the FBL cell in the incubator, the used Digestive system of passage contains 0.02%EDTA 0.25% pancreatin.
2.2 virus culture
FBL cell (BIV-R29/FBL) and normal 1: 1 mixed culture of FBL cell with BIV (R29) infects increase BIV in a large number.
2.3 medicine pair cell poison experiment (FBL)
Density with 40000/ hole is laid on 24 orifice plates with the FBL cell, and cell density increases to 50000/ hole after 24 hours, adds the medicine of 0.1mg/ml, 0.15mg/ml, 0.2mg/ml, with the negative contrast of normal cell, and the positive contrast of AZT.(about 72 hours) counted every porocyte number after normal cell covered with, and determined medicine 50% lethal concentration (IC
50)
2.4 infecting BIV, medicine causes that synplasm forms the inhibition experiment
Density with 40000/ hole is laid on 24 orifice plates with the FBL cell, and the density with 2000/ hole after 24 hours adds BIV-R29/FBL (MOI=1: 25), add medicine (concentration＜IC to be measured simultaneously
50), with the positive contrast of AZT, the negative contrast of equal-volume PBs, the counting synplasm forms quantity after 72 hours.
Tremella polysaccharide is to the cellulotoxic experiment of tire ox pneumonocyte
| Sample | Concentration (mg/ml) | The survivaling cell number | ||
| Hole 1 | Hole 2 | Hole 3 | ||
| Culture medium A ZT normal cell tremella polysaccharide | / 0.0026 / 0.2 0.1 0.05 | 0 5.8 6.5 5.6 6.8 5.8 | 0 5.9 6.0 5.5 6.3 5.9 | 0 5.3 5.6 4.9 5.1 6.2 |
Cell toxicant result: tremella polysaccharide IC
50＞0.2mg/ml
Table 3 tremella polysaccharide infects the restraining effect of the synplasm formation that causes to BIV
| Sample | Concentration (mg/ml) | Synplasm quantity | ||
| Hole 1 | Hole 2 | Hole 3 | ||
| Culture medium A ZT BIV-R29 | / 0.0026 / | - - ++++ | - + ++++ | - - ++++ |
| The normal cell tremella polysaccharide | / 0.2 0.2 0.15 | - +++ +++ ++++ | - ++ ++ +++ | - +++ ++ ++++ |
Synplasm number and control wells are close ++ ++ the synplasm number lacks 75% than control wells +++
The synplasm number than control wells lack 50%++ synplasm number than control wells lack 25%+
Substantially not having synplasm occurs
Above-mentioned pharmacological evaluation shows:
(1) toxicity test shows, tremella polysaccharide toxicity is lower, its IC
50All greater than 0.2mg/ml.
(2) synplasm that is caused by bovine immunodeficiency virus suppresses description of test, the effect that tremella polysaccharide has certain inhibition synplasm to form when the concentration of 0.2mg/ml.
Seven, antioxygenation:
At H
2O
2In the test of the erythrocyte hemolysis that causes, with the distilled water hemolysis rate be 100%, inhibiting rate is 0% to calculate, ATF-A, ATF-B, ATF-C, ATF-D inhibiting rate are respectively 78.6%, 68.8%, 71.5%, 74.4%;
At polysaccharide to ultra-oxygen anion free radical (O
2-.) scavenging(action) experiment in, ATF-A, ATF-B, ATF-C, the highest inhibiting rate of ATF-D are respectively 21.4%, 28.4%, 36.6%, 53.0%;
In removing the hydroxyl radical free radical experiment, the EC of ATF-A, ATF-B, ATF-C, ATF-D
50Be respectively 233.6mg/L, 603mg/L, 191mg/L, 195mg/L.
Homogeneous tremella polysaccharide of the present invention and extracting method thereof are compared with prior art has following advantage:
1. homogeneous tremella polysaccharide of the present invention has the content height, good stability, and effective constituent is definite, the characteristics that quality is easy to control.
2. utilize tremella spore saleratus, extract tremella polysaccharide and be not subjected to place, time limitation, can reduce cost, be easy to large-scale industrial production.
3. equal one polysaccharide molecular weight of the present invention is less, good water solubility, and the purity height not only can be made into oral preparations but also be easy to make liquid preparation, thereby makes effective constituent better bring into play curative effect, to satisfy requirement of different patients.
Embodiment:
The present invention is described further below by embodiment, but be not to be limitation of the present invention:
Embodiment 1
(1) gets 150g tremella spore saleratus, use the 3000ml poach, about 5 hours.After the cooling room temperature, extracting solution is centrifugal, and the residue after the extraction is used poach again, and the cooling back is centrifugal, merges supernatant liquor twice, is concentrated into little thickness with Rotary Evaporators.Add triplication ethanol, leave standstill the back suction filtration and get precipitation.Precipitation is washed with a little ether, and vacuum-drying gets product 80g, is Crude polysaccharides.
(2) get 10 gram Crude polysaccharides, add the 500ml aquae destillata, heating for dissolving, after the cooling, inclining supernatant liquor, and insolubles adds 100ml water heating for dissolving again.After the cooling, centrifugal, get supernatant liquor and last supernatant liquor and merge, amount to supernatant liquor 600ml, add 125ml chloroform and 25ml propyl carbinol, mechanical stirring 20 minutes produces milky white precipitate.Centrifugal, get supernatant liquor and add chloroform and propyl carbinol again, restir, centrifugal, so repeatedly till do not have a proteins react.To an amount of, with aquae destillata dialysis three days, solution added the triplication ethanol sedimentation in the dialysis tubing with supernatant concentration.Suction filtration is washed precipitation with dehydrated alcohol, acetone, ether.Put drying in the vacuum drier, get fine silver fungus polysaccharides 5.18g.
(3) get the fine silver fungus polysaccharides of 500mg,, at capital, carry out column chromatography with DEAE-SephadexA-25 with supernatant liquor with 6ml 0.1M NaCl dissolving.Behind the sample introduction, with 0.1M NaCl wash-out, fraction collection is with 752 spectrophotometric determination absorbancys.The reaction that always is eluted to polysaccharide is negative.Obtain two kinds of homogeneous tremella polysaccharides.Called after WTF-A and WTF-B.
Embodiment 2
(1) water extraction: get 300g tremella spore fermented product, add the water of 40 times of amounts, boil and carry about 4 hours, keep the consistence of the water yield in the boiling part.Put behind the poach to room temperature, extracting solution is centrifugal, and the residue after the extraction adds about 40 times water, boils with identical method and carries twice and centrifugation collecting precipitation.The gained precipitation places 50 ℃ of baking ovens to dry, and weighs, and weight is 188.8g.Precipitation is Crude polysaccharides.
(2) alkali lye extracts: get the 140g Crude polysaccharides, add 0.5mol/LNaOH 1000ml, extracted 6 hours, and centrifugal.Precipitation extracts twice again with identical method, merges the supernatant liquor of three gained, and precipitation is put in addition.Supernatant liquor is evaporated to 1/3rd of about original volume, is neutralized to PH=7, neutralizer is evaporated to small volume with the HCl of 1mol/L.Neutralizer after concentrating was dialysed three days with flowing water.Solution after the dialysis is amassed 85% ethanol precipitating with triploid, and centrifugal back collecting precipitation is 0.5mol/LNaOH and extracts the alkali-extracted polysaccharide that obtains.
(3) refining polysaccharide: the 0.5mol/LNaOH extract is added about 700ml water stirring and dissolving, add 175mlSevag reagent (chloroform: propyl carbinol=4: 1), after the mechanical stirring 0.5 hour, centrifugal the gained supernatant liquor is removed albumen with this method repeatedly, till detecting no proteins react with ninhydrin reagent.The gained supernatant liquor is evaporated to small volume uses flowing water dialysis three days, solution amasss 85% ethanol precipitating with triploid in the dialysis tubing.Suction filtration, precipitation places vacuum drier dry with dehydrated alcohol, acetone, ether washing, gets 0.5mol/LNaOH and extracts the refining polysaccharide of gained, is weighed as 4.470g.
(4) homogeneous tremella polysaccharide: get 600mg 0.5mol/LNaOH and extract DEAE-32-cellulose chromatography column on the refining polysaccharide of gained, use the 0.1mol/LNaCl eluant solution, fraction collection obtains the 112mg homogeneous tremella polysaccharide, called after " ATF-A ".
Embodiment 3
Under aseptic condition, get homogeneous tremella polysaccharide 18g, place container, add injection water 1000ml, add N.F,USP MANNITOL 50g, lactose 20g, carry out the autoclave sterilization sterilization by the requirement of injection, add the 1.5g activated carbon, adopt filtering with microporous membrane, filtrate is carried out packing by every 0.6ml, adopts freeze-drying, make yellow loose block, seal promptly.
Embodiment 4
Get homogeneous tremella polysaccharide 100g under the aseptic condition, join among the water for injection 24000ml, add 60g Sulfothiorine and stirred 30 minutes, add the 30g gac, whip attachment 30 minutes, carbon removal, smart filter, embedding, sterilization promptly get injection liquid.
Or get homogeneous tremella polysaccharide 100g under the aseptic condition, and add among the water for injection 24000ml, after the G4 funnel filtered, the packing sterilization made injection liquid.
Embodiment 5:
Homogeneous tremella polysaccharide 1000g, medical starch 600g, dextrin 600g, 50% ethanol is an amount of, above-mentioned raw materials is fully mixed make particle, at 60-70 ℃ of dry 2-4 hour, makes 1000, makes every to contain homogeneous tremella polysaccharide 0.3g.
Embodiment 6:
Homogeneous tremella polysaccharide 1000g, medical starch 1000g, 50% ethanol is an amount of, granulates, whole grain, oven dry, dress 1# capsule, every 0.2g
Embodiment 7:
Homogeneous tremella polysaccharide 20g, polyoxyethylene glycol-4000 10g, polyoxyethylene glycol-6000 30g, Sodium Pyrosulfite is an amount of, and heating and melting temperature (40-90 ℃) makes raw material and auxiliary material fully miscible, drip and make 1000 dripping pills, make every dripping pill contain homogeneous tremella polysaccharide 20mg.
Although the present invention has done detailed description in conjunction with its special embodiment, clearly concerning the skilled people in present technique field, still can make various changes and improvements, can not depart from spirit of the present invention and protection domain.
Claims (9)
1, multiple homogeneous tremella polysaccharide, it be by the tremella spore fermented product through poach, alcohol precipitation, remove albumen or NaOH aqueous solution gradient extraction with different concns after, obtain content at the homogeneous tremella polysaccharide more than 90% through column chromatography for separation, purifying; Mainly contain following composition:
Alkali desilver fungus polysaccharides ATF-A, ATF-B, ATF-C, ATF-D;
Water desilver fungus polysaccharides WTF-A, WTF-B;
Their weight-average molecular weight of high effective liquid chromatography for measuring is respectively 62830,63710,58962,51454,73000 and 68000; Wherein,
ATF-A mainly is made up of D-glucose, D-wood sugar, D-seminose;
ATF-B mainly is made up of D-glucose, D-wood sugar, D-seminose;
ATF-C mainly is made up of D-glucose, L-rhamnosyl;
ATF-D mainly is made up of D-glucose, D-wood sugar, D-seminose, D-semi-lactosi; WTF-A mainly is made up of L-Fucose, D-wood sugar, D-seminose, D-semi-lactosi and D-glucose; WTF-B mainly is made up of L-Fucose, D-wood sugar, D-seminose, D-glucose.
2, the extracting method of the described multiple homogeneous tremella polysaccharide of claim 1 comprises the steps:
The extraction of A, water-soluble homogeneous tremella polysaccharide:
(1) tremella spore saleratus poach is carried, cooling, centrifugal, the supernatant concentration alcohol precipitation filters, the dry Crude polysaccharides that gets;
(2) Crude polysaccharides removes albumen through the sevage method, and with the flowing water dialysis, 70-90% ethanol precipitating must be made with extra care polysaccharide;
(3) refining polysaccharide with 0.1-5mol/L sodium chloride aqueous solution wash-out, is collected different elutriants through column chromatography for separation, concentrates back alcohol precipitation drying, makes water and carries homogeneous tremella polysaccharide, WTF-A, WTF-B;
The extraction of B, slightly water-soluble homogeneous tremella polysaccharide:
(1) after going out water-soluble component with hot water extraction, with 0.1mol/L-10mol/LNaOH aqueous solution gradient extraction, hydrochloric acid is neutralized to neutrality, is concentrated into small volume to remaining slightly water-soluble component, and with the flowing water dialysis, 70-90% ethanol precipitating makes alkali-extracted polysaccharide;
(2) alkali is carried Crude polysaccharides and is removed albumen through the sevage method, and with the flowing water dialysis, 70-90% ethanol precipitating must be made with extra care polysaccharide;
(3) refining polysaccharide with 0.1mol/L-5mol/L sodium chloride aqueous solution wash-out, is collected different elutriants through column chromatography for separation, concentrates the back alcohol precipitation, and vacuum-drying makes alkali and carries homogeneous tremella polysaccharide, ATF-A, ATF-B, ATF-C, ATF-D.
3, as the extracting method of homogeneous tremella polysaccharide as described in the claim 2, chromatography column wherein comprises DEAE-32-cellulose, DEAE-52-cellulose, DEAE-Sephadex gel A-25 and Sephadex G-200, Sephadex G-100, Sephadex G-150.
4, a kind of is the pharmaceutical composition of activeconstituents with the homogeneous tremella polysaccharide, and said composition contains the homogeneous tremella polysaccharide for the treatment of significant quantity, and one or more pharmaceutically acceptable carrier.
5, pharmaceutical composition as claimed in claim 4, wherein, the treatment significant quantity of homogeneous tremella polysaccharide is 0.1-500mg.
6, pharmaceutical composition as claimed in claim 4, wherein, the treatment significant quantity of homogeneous tremella polysaccharide is 1-200mg.
7, as the described pharmaceutical composition of claim 4-6, wherein, said composition can oral preparations, the form of liquid preparation or external preparation exists.
8, pharmaceutical composition as claimed in claim 7, wherein, oral preparations is tablet, capsule, slow releasing tablet, dripping pill, granule or oral liquid; Liquid preparation is freeze-dried powder, injection liquid, large and small transfusion or intravenous drip liquid; External preparation is creme or paste.
9, homogeneous tremella polysaccharide is in the application that is used for preparing enhancing immunity, antitumor, anti-radiation, leukocyte increasing, antiviral, antiaging agent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410072865 CN1778824A (en) | 2004-11-25 | 2004-11-25 | Multiple homogeneous tremella polysaccharide, its extraction and medicinal composition with this compound as active components |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410072865 CN1778824A (en) | 2004-11-25 | 2004-11-25 | Multiple homogeneous tremella polysaccharide, its extraction and medicinal composition with this compound as active components |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1778824A true CN1778824A (en) | 2006-05-31 |
Family
ID=36769288
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200410072865 Pending CN1778824A (en) | 2004-11-25 | 2004-11-25 | Multiple homogeneous tremella polysaccharide, its extraction and medicinal composition with this compound as active components |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1778824A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101086004B (en) * | 2007-06-21 | 2010-05-19 | 侯建明 | Enzyme method reverse extraction process for white fungus polysaccharidase |
| CN102336840A (en) * | 2011-10-17 | 2012-02-01 | 浙江大学 | Triple-helical Tremellan, preparation method and application thereof |
| CN102746418A (en) * | 2012-08-01 | 2012-10-24 | 吉林农业大学 | Sansevieria trifasciata polysaccharides as well as preparation method and application thereof |
| CN103006460A (en) * | 2012-12-31 | 2013-04-03 | 涂桂洪 | Skin-caring composition and preparation method and applications thereof |
| CN106074214A (en) * | 2016-07-13 | 2016-11-09 | 袁春华 | A kind of preparation method of the anti-aging facial film of radioprotective |
| CN107011451A (en) * | 2016-04-25 | 2017-08-04 | 上海辉文生物技术股份有限公司 | White fungus different poly- polysaccharide and its extract, preparation method and application |
| CN111991360A (en) * | 2020-10-27 | 2020-11-27 | 中科院大连化学物理研究所张家港产业技术研究院有限公司 | Tremella polysaccharide oral tablet and preparation process thereof |
| CN112316133A (en) * | 2020-11-16 | 2021-02-05 | 高其品 | Preparation method and application of tremella polysaccharide immunologic adjuvant for injection |
| CN113974158A (en) * | 2021-10-15 | 2022-01-28 | 江西仙客来生物科技有限公司 | Ganoderma lucidum, agaricus blazei murill and epimedium oral liquid |
| CN116574197A (en) * | 2023-05-12 | 2023-08-11 | 广西壮族自治区农业科学院 | Tremella polysaccharide and application thereof |
-
2004
- 2004-11-25 CN CN 200410072865 patent/CN1778824A/en active Pending
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101086004B (en) * | 2007-06-21 | 2010-05-19 | 侯建明 | Enzyme method reverse extraction process for white fungus polysaccharidase |
| CN102336840A (en) * | 2011-10-17 | 2012-02-01 | 浙江大学 | Triple-helical Tremellan, preparation method and application thereof |
| CN102336840B (en) * | 2011-10-17 | 2012-11-21 | 浙江大学 | Triple-helical Tremellan, preparation method and application thereof |
| CN102746418A (en) * | 2012-08-01 | 2012-10-24 | 吉林农业大学 | Sansevieria trifasciata polysaccharides as well as preparation method and application thereof |
| CN103006460A (en) * | 2012-12-31 | 2013-04-03 | 涂桂洪 | Skin-caring composition and preparation method and applications thereof |
| CN107011451A (en) * | 2016-04-25 | 2017-08-04 | 上海辉文生物技术股份有限公司 | White fungus different poly- polysaccharide and its extract, preparation method and application |
| CN107011451B (en) * | 2016-04-25 | 2019-11-08 | 上海辉文生物技术股份有限公司 | The different poly- polysaccharide of tremella and its extract, preparation method and application |
| CN106074214A (en) * | 2016-07-13 | 2016-11-09 | 袁春华 | A kind of preparation method of the anti-aging facial film of radioprotective |
| CN111991360A (en) * | 2020-10-27 | 2020-11-27 | 中科院大连化学物理研究所张家港产业技术研究院有限公司 | Tremella polysaccharide oral tablet and preparation process thereof |
| CN112316133A (en) * | 2020-11-16 | 2021-02-05 | 高其品 | Preparation method and application of tremella polysaccharide immunologic adjuvant for injection |
| CN112316133B (en) * | 2020-11-16 | 2024-02-27 | 高其品 | Preparation method and application of tremella polysaccharide immunoadjuvant for injection |
| CN113974158A (en) * | 2021-10-15 | 2022-01-28 | 江西仙客来生物科技有限公司 | Ganoderma lucidum, agaricus blazei murill and epimedium oral liquid |
| CN116574197A (en) * | 2023-05-12 | 2023-08-11 | 广西壮族自治区农业科学院 | Tremella polysaccharide and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1872282A (en) | Active extractive of Foliumnerviliae, preparation method and application | |
| CN101249259B (en) | High content and high activity oral polysaccharide-peptide and preparing method and application of the same | |
| CN1596118A (en) | Therapeutical treatments | |
| CN1772081A (en) | Diarrhea treating chinese medicine composition | |
| CN1778824A (en) | Multiple homogeneous tremella polysaccharide, its extraction and medicinal composition with this compound as active components | |
| CN1762967A (en) | Enoxolone derivative, preparation method and uses | |
| CN1723955A (en) | Extractive of rhizome belamcandae, prepn. method and use thereof | |
| CN1557842A (en) | Process for extracting polygonatum polysaccharides, preparing process for medical preparation and use thereof | |
| CN1118471C (en) | Medicine containing tan matter caesalpinia extract | |
| CN101032532A (en) | Medicine composition including active substrates extracted from Sang Huang, the preparing method and the application in the producing of medicine thereof | |
| CN1323668C (en) | Pharmaceutical compositions and its application | |
| CN109602759A (en) | The purposes of kusamaki broad-leaved podocarpus seed and receptacle polysaccharide | |
| CN101040890A (en) | Compound of sulfoacid flavonecosid component in chickweed and the antivirus application and the preparing method | |
| CN100584345C (en) | Distillage of Ardisia chinensis Benth of possessing function of antivirus, extraction method and application | |
| CN1493578A (en) | Preparation method of golden peach glycoside and its new use in medicine | |
| CN1116891C (en) | Hepatitis B treating medicine | |
| CN1184229C (en) | Furost saponine analogue and its separation process and use | |
| CN1201745C (en) | Medicine combination contg. effective part of astragalus root | |
| CN1566136A (en) | Pasqueflower notoginsenosides and extraction method, medicinal uses and pharmaceutical preparation thereof | |
| CN1253175C (en) | Medicinal composition, its preparation method and use | |
| CN1919270A (en) | Composition, exract, and pharmaceutical use thereof | |
| CN1903308A (en) | Active position of Omithogalum caudatum Ait its prepn. method, medicine and application thereof | |
| CN108542940A (en) | A kind of Chinese medicine composition prepare prevent and/or treatment hand-foot-and-mouth disease drug in application | |
| CN1256119C (en) | Traditional Chinese medicine preparation for treating nonspecific colonitis and its preparing process | |
| CN1943584A (en) | Medicinal composition for treating cardio-cerebral vascular disease and its preparing method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |