CN104055803A - Fomes fomentarius compound extract and preparation method and application thereof - Google Patents
Fomes fomentarius compound extract and preparation method and application thereof Download PDFInfo
- Publication number
- CN104055803A CN104055803A CN201210355671.0A CN201210355671A CN104055803A CN 104055803 A CN104055803 A CN 104055803A CN 201210355671 A CN201210355671 A CN 201210355671A CN 104055803 A CN104055803 A CN 104055803A
- Authority
- CN
- China
- Prior art keywords
- extract
- fomes fomentarius
- compound
- cell
- tumor
- 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
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to the technical field of biological medicine. In particular, the invention relates to a fomes fomentarius compound extract, a preparation method and an application thereof. The fomes fomentarius compound extract is prepared from the following components according to a weight ratio of 1-10 parts of fomes fomentarius:1-9 parts of phellinus pini:1 part of ganoderma lucidum; the extraction method of the compound extract comprises: preparing a fomes fomentarius compound, mixing, crushing, adding pure water, performing water extraction and alcohol precipitation to obtain a supernatant, performing reduced pressure concentration, and performing vacuum drying of the concentrate to obtain the product. The content of triterpene and triterpene saponin in the fomes fomentarius compound extract of the invention is not less than 5.5%. The extract not only has significant in-vivo and in-vitro antineoplastic efficacy, but also has no obvious acute toxicity; the maximum tolerance dose for mice is 0.2 g (equivalent to 4.58 g of crude drugs), which is 327 times of human oral dosage. The extract can be prepared into compound preparation when mixed with pharmaceutically acceptable carriers, and the compound preparation can be oral liquid, injection, suspension, pills, tablets, capsules, powder, granules, or drop pills.
Description
1 technical field
The present invention relates to biological medicine technology field.Specifically, the present invention relates to a kind of Fomes Fomentarius compound extract and preparation method thereof, this compound extract is mainly used in the growth of in vitro and in vivo inhibition tumor cell.
2 background technologies
In recent years, China's tumor incidence is in the fast rise phase, and annual pathogenesis of cancer number approximately 2,600,000 is dead 1,800,000, has become Chinese city and urban residents' first cause of the death.Because total surgical resection, chemotherapy and radiation all exist inherent shortcoming, natural drug or Traditional Chinese Medicine Anti tumor research are by pay attention to day by day.Medicinal fungi is an important component part in natural drug, and one of antitumor action effect that to be medicinal fungi important.
Wherein, Fomes fomentarius Fomes fomentarius (L.:Fr) Kick. has another name called Pyropolyporus fomentarius (L.ex Fr.) Teng, is the medicinal fungi that Polyporaceae, shelf fungus belong to.In " China's book on Chinese herbal medicine ", record: Fomes fomentarius sporophore stockless.The cap shape of a hoof, major diameter 3-40cm, minor axis 2-27cm, thick 2-18cm.There is thick cutin cot on surface, and Lycoperdon polymorphum Vitt, light brown or black are smooth, have obvious concentric ring rib.Light brown or the Lycoperdon polymorphum Vitt of mouth of pipe face, mouth of pipe circle, between every 1mm 3-4.The visible tube multilamellar in vertical section, every bed thickness 5-25mm.Suberin.Feeble QI, lightly seasoned.Its pharmacological action is mainly: showing the tolerance that improves experiment mice hypobaric hypoxia, extending its time-to-live, having the effect of resisting fatigue, high temperature resistance.And flow velocity, the fluidised form that can affect anoxia body Mesentery microcirculation, have improvement effect to microcirculation.Cure mainly: removing food stagnancy; Blood stasis dispelling; Anticancer (the esophageal carcinoma; Gastric cancer and uterus carcinoma).Prove by mouse experiment: in Fomes fomentarius, isolated tannins phenol thing has obvious killing action to mice transplanted tumor S180, and the further investigations such as Zhang Liping find that Fomes fomentarius polysaccharide there is no direct killing effect to S180 tumor cell, but by activating the immunologic function of animal body, the activity of activating macrophage, natural killer cell, killer T cell, thus strengthen tumour immunity function.
Phellin uspini Phellinus pini. has another name called Folium Pini Ganoderma, Phellinus pini, for Polyporaceae, shelf fungus belong to medicinal fungi." Macrofungi From China primary colors illustrated handbook " recorded: its sporophore is perennial, suberin, and nothing, the flat rice of cap is spherical to the shape of a hoof, 4-14 centimetre 7 × 24 centimetres, seldom reaches 40 centimetres of 23 cm x, thick 2.5-18.5 centimetre or larger.There is the gluey cot of bronzing at the capping initial stage, and gradually keratinization and grey be to black, and have wide rib band, and edge is toward blunt, initial stage near-white, and rear gradual change is faint yellow to red maroon, and downside is without hymenium.Bacterial context near-white, to khaki, has ring grain, and suberin is to wooden.Thick 0.5-2 centimetre, bitter in the mouth.Tube multilamellar, every bed thickness 3-5 millimeter, faint yellow, mouth of pipe white is to milky.Spore oval or ellipse, colourless.Be born on the acerose trunks such as pine, PiceameyeriRehd. Et Wils., fir, Chinese hemlock spruce and larch or rotten wood.Also be born in once in a while on the withered rotten wood of broad leaf tree.Wang Wenhang, Liu Anjun etc. compare systematic research to the pharmacological action of phellin uspini, result shows: three kinds of polysaccharide of Folium Pini Ganoderma have all improved SOD in mice serum, the heart, liver, brain, spleen, GSH-Px vigor to some extent, reduce MDA growing amount, and less on the impact of TOAC value; Meanwhile, the three benefit sugar serum NO level growing amount in Mice Body that has been significantly increased.Meanwhile, they propose the Antioxidation Mechanism of three kinds of polysaccharide may be different from other antioxidant, and it is dual increase effect to oxidative pressure in Mice Body and antioxygen level, may be because polysaccharide causes that as immune material the immunoreation of body causes.Phellin uspini be red wild pine in Coniferous forest, larch, PiceameyeriRehd. Et Wils. topmost, endanger maximum fiery root and butt rot, be undoubtedly a kind of cancer for Coniferous forest, but be but for the mankind potion good medicine that suppresses tumor, treatment cancer.In " Macrofungi From China primary colors illustrated handbook " 73 pages, record: Folium Pini sporophore, containing eburicoic acid isoreactivity material, is in all funguses, to treat the best kind of cancer effect, and fruiting body extract all reaches 100% to the suppression ratio of white mice sarcoma 180 and ehrlich carcinoma.But Folium Pini has toxicity, for reaching the object of strengthening vital QI to eliminate pathogenic factors in theory of Chinese medical science, so must coordinating with other Chinese medicine, Folium Pini just can remove toxicity, safe handling.Adopting phellin uspini is at present the mechanism that Main Ingredients and Appearance is treated effective formula of all kinds of cancers, only has two both at home and abroad, the CELG of a Jia Shi U.S., the Chinese grass luxuriant growth that another domestic are domestic.
Ganoderma lucidum (Leyss. Ex Fr.) Karst. Ganoderma lucidum (Leyss.ex Fr) Karst is Polyporaceae, Ganoderma medicinal fungi.Record: Ganoderma lucidum (Leyss. Ex Fr.) Karst. profile is umbrella cap kidney shape, semicircle or subcircular, diameter 10～18cm, thick 1～2cm according to " Chinese Pharmacopoeia ".Cot is hard, and yellowish-brown is to bronzing, glossy, tool ring-type rib and radial wrinkle, and thin edge and truncate, often slightly involute.Bacterial context white is to light brown.Stem is cylindrical, adnation, few wilfully, long 7～15cm, diameter 1～3.5cm, bronzing is to puce, light.Spore is tiny, yellowish-brown.Feeble QI perfume (or spice), bitter and puckery flavor.Its main effect is: invigorating QI and tranquilization, relieving cough and asthma.For dizzy sleeplessness, shortness of breath and palpitation, asthenia cough with asthma.In addition, Ganoderma lucidum (Leyss. Ex Fr.) Karst. antitumor efficacy is by wide coverage, and its antitumor mechanism is also furtherd investigate, mainly comprise: 1. short apoptosis: by promoting Bax to express, suppress the gene expression such as Bcl-2 and Bcl-xl simultaneously, thereby suppress the growths such as lymphoma, inflammatory breast cancer, impel its apoptosis; 2. growth inhibited: by suppressing cyclin cyclinD1, suppress ovarian cancer cell growth; 3. improve immunity: ganoderan and triterpene substance can extensively regulate the function of immunocyte and immunity-associated cell, comprise B cell, T cell, NK cell, dendritic cell, macrophage etc., simultaneously they also time to promote TNF-alpha expression, reduce IL-10, TGF-β 1 and vegf expression, strengthen immune response strength.
At present, Chinese medicine, natural drug compound recipe have become one of the main form of medication of curing the disease, and the Fungal community composition compound recipe of different pharmacological mechanism is more conducive to antineoplaston.This research proposes Fomes fomentarius, phellin uspini and Ganoderma lucidum (Leyss. Ex Fr.) Karst. triplicity first, to reach righting gas, improves the effect of tumour immunity, tumor-killing.
3 summary of the invention
The object of the invention is to provide a kind of Fomes Fomentarius compound extract and preparation method of high-efficiency low-toxicity.
Another object of the present invention is to provide this extract in vivo with extracorporeal anti-tumor in purposes.This extract remarkable killing tumor cell in vitro, and effectively suppress mouse interior tumor growth; And mice is oral and lumbar injection is not all found overt toxicity, and maximum tolerated dose is 10g/kg.
The present invention relates to a profit and have Fomes Fomentarius compound extract remarkable antitumor efficacy, hypotoxic and preparation method.
In Fomes Fomentarius compound recipe of the present invention, Fomes fomentarius: phellin uspini: the weight ratio of Ganoderma lucidum (Leyss. Ex Fr.) Karst. is 1～10: 1～9: 1.
Preferably, in Fomes Fomentarius compound recipe of the present invention, Fomes fomentarius: phellin uspini: the weight ratio of Ganoderma lucidum (Leyss. Ex Fr.) Karst. is 2～7: 1～6: 1.
More preferably, in Fomes Fomentarius compound recipe of the present invention, Fomes fomentarius: phellin uspini: the weight ratio of Ganoderma lucidum (Leyss. Ex Fr.) Karst. is 4～5: 1～3: 1.
It is as follows that the present invention prepares Fomes Fomentarius compound extract method step:
Step 1: Fomes fomentarius, phellin uspini and Ganoderma lucidum (Leyss. Ex Fr.) Karst. require to be made into Fomes Fomentarius compound recipe according to weight ratio, pulverizes after mixing;
Step 2: the Fomes Fomentarius compound recipe after pulverizing adds pure water, 100 DEG C of heating and refluxing extraction 3 times, each 2 hours, solid-liquid ratio was respectively 1: 20,1: 15 and 1: 10, and 4 DEG C left standstill after 24 hours, 9000rmin
-1centrifugal 30 minutes, 20 times of volumes of supernatant concentrating under reduced pressure.Afterwards, concentrated solution adds 95% ethanol again, and making ethanol final concentration is 80%, leaves standstill after 24 hours 9000rmin at 4 DEG C
-1centrifugal 30 minutes, get supernatant, 20 times of volumes of concentrating under reduced pressure, concentrated solution vacuum drying, to obtain final product.
Fomes Fomentarius compound extract prepared by the present invention, its component analysis result shows that it contains organic acid, aminoacid, peptide and protein, sugar, polysaccharide and glycoside, saponin, lactone, coumarin and glycoside thereof, the Multiple components such as plant sterol and Triterpenoid and volatilization oil and grease, wherein the content of triterpenes and triterpene saponin is not less than 5.5%.
Fomes Fomentarius compound extract prepared by the present invention, its purposes is: (1) this extract has remarkable lethal effect to tumor cell lines such as human lung adenocarcinoma cell (A549), human cervical carcinoma cell (Hela), human liver cancer cell (QGY), Human Prostate Cancer Cells (DU145), human colon cancer cell (HCT116), human esophagus cancer cell (Eca109), human breast cancer cell (MDA-MB-435S), gastric carcinoma cells (SGC-7901), human osteosarcoma cell cell (Saos-2), human osteosarcoma cells (U2Os) in vitro.(2) effectively cell death inducing of this extract, after final concentration is 100 μ g/ml effect 24h, the apoptosis rate of A549 is 14.8%.(3) this extract has significant inhibitory action to murine sarcoma S180 and Lewis lung cancer transplanted tumor model tumor in vivo, in the time that dosage is 150mg/kg, is 57.7% to the tumour inhibiting rate of S180 sarcoma; In the time that dosage is 100mg/kg, be 56.5% to the tumour inhibiting rate of Lewis lung cancer.(4) this extract can effectively improve tumor-bearing mice immunologic function, can make the thymus index of tumor-bearing mice and peripheral blood TNF-a content improve.
Fomes Fomentarius compound extract prepared by the present invention mixes with pharmaceutically acceptable carrier, can be made into compound preparation, and wherein compound preparation comprises oral liquid, injection, suspensoid, pill, tablet, capsule, powder, granule or drop pill.
Compared with the prior art the present invention, has following marked feature:
(1) provide a kind of new antineoplastic Fomes Fomentarius compound recipe: Fomes fomentarius antineoplastic action mechanism is its toxic action, as monarch drug in compound recipe, and phellin uspini also has remarkable antitumor action, as ministerial drug, and Ganoderma lucidum (Leyss. Ex Fr.) Karst. is improved effect of immunity of organisms, with the above two compatibility uses, there is the effect of fine strengthening vital QI to eliminate pathogenic factors.
(2) provide a kind of new antitumor Fomes Fomentarius compound extract, the extracting method of this extract is: water extract-alcohol precipitation, and then centrifuging and taking supernatant, concentrated, dry, to obtain final product.
(3) this extract component analysis result shows that it contains organic acid, aminoacid, peptide and protein, sugar, polysaccharide and glycoside, saponin, lactone, coumarin and glycoside thereof, the Multiple components such as plant sterol and Triterpenoid and volatilization oil and grease, wherein the content of triterpenes and triterpene saponin is not less than 5.5%.
(4) effect that this extract has in obvious body, extracorporeal suppression tumor cell is grown; Can improve tumor-bearing mice immunologic function, can make the thymus index of tumor-bearing mice and peripheral blood TNF-a content improve; And this extract mice is oral and lumbar injection does not have overt toxicity, and maximum tolerated dose is 10g/kg.
(5) this extract is soluble in water, and aqueous solution color is brown.
(6) this extract mixes with pharmaceutically acceptable carrier, can be made into compound preparation
4 brief description of the drawings
Fig. 1 Fomes Fomentarius compound extract is on the apoptotic impact of A549
5 detailed description of the invention
Below in conjunction with specific embodiment, further illustrate the present invention.Should be understood that these embodiment are only not used in and limit the scope of the invention for the present invention is described.The experimental technique of unreceipted actual conditions in the following example, conventionally according to normal condition.
The preparation of embodiment 1 Fomes Fomentarius compound extract
Fomes fomentarius 60g, phellin uspini 30g and Ganoderma lucidum (Leyss. Ex Fr.) Karst. 15g, pulverize, and adds water, 100 DEG C of heating and refluxing extraction 3 times, each 2h, solid-liquid ratio is respectively 1: 20,1: 15 and 1: 10, filter merging filtrate, 4 DEG C of standing 24h, the centrifugal 30min of 9000r/min, the 20 times of volumes of liquid concentrating under reduced pressure that above disappear, concentrated solution adds 95% ethanol, making ethanol final concentration is 80%, 4 DEG C of standing 24h, the centrifugal 30min of 9000r/min, 20 times of volumes of supernatant concentrating under reduced pressure, concentrated solution vacuum drying, to obtain final product.
The preparation of embodiment 2 Fomes Fomentarius compound extracts
Fomes fomentarius 70g, phellin uspini 20g and Ganoderma lucidum (Leyss. Ex Fr.) Karst. 15g, pulverize, and adds water, 100 DEG C of heating and refluxing extraction 3 times, each 2h, solid-liquid ratio is respectively 1: 20,1: 15 and 1: 10, filter merging filtrate, 4 DEG C of standing 24h, the centrifugal 30min of 9000r/min, 20 times of volumes of supernatant concentrating under reduced pressure, concentrated solution adds 95% ethanol, making ethanol final concentration is 80%, 4 DEG C of standing 24h, the centrifugal 30min of 9000r/min, 20 times of volumes of supernatant concentrating under reduced pressure, concentrated solution vacuum drying, to obtain final product.
Embodiment 3 Fomes Fomentarius compound extract component analyses
(1) component analysis
Fomes Fomentarius compound extract is dissolved in to 80% alcoholic solution, obtains test liquid, then adopt system trial test to differentiate it.1. test tube trial test adds related reagent to filling in the test tube of test liquid, judges the existence of certain chemical composition according to reacting phenomenons such as color, precipitation, fluorescence.2. circular filter paper layer method drawn test liquid point sample on circular filter paper with capillary microcap, after dry, makes ethanol from center deployment, and spray, with different developers, judges the existence of certain chemical composition according to phenomenons such as the fluorescence of the color, speckle or the generation that occur.Result is as shown in table 1, and known its contains organic acid, sugar, polysaccharide and glycoside, saponin, lactone, coumarin and glycoside thereof, plant sterol, Triterpenoid and volatilization oil and grease.May contain alkaloid, aminoacid, peptide and protein, pyrite and glycoside thereof and anthraquinone and glycoside thereof.
Table 1 Fomes Fomentarius compound extract chemical composition
Note：“+”means positive reaction；“-”means negative reaction
(2) total triterpene and triterpene saponin assay
1. the preparation of reference substance solution
Precision takes ursolic acid reference substance 10.7mg, is placed in 50mL volumetric flask, adds methanol and makes it to dissolve and be diluted to scale, shakes up, and to obtain final product.
2. the preparation of need testing solution
Precision takes compound extract 200mg, is placed in 200mL volumetric flask, adds dissolve with methanol and is diluted to scale, shakes up, and to obtain final product.
3. the preparation of standard curve
The accurate ursolic acid solution 0,0.1,0.2 of drawing, 0.3,0.4,0.5,0.6mL is placed in respectively tool plug test tube, add and fling to solvent, 5% vanillin-glacial acetic acid solution the 0.2mL and the perchloric acid 1.2mL that add new preparation heat 15min in 70 DEG C of waters bath with thermostatic control, and flowing water is chilled to room temperature, adding ethyl acetate to make total amount is that 5mL. shakes up, and surveys absorbance after 15min at 550nm place.Taking reagent blank as reference liquid, drawing standard curve.Obtain regression equation y=4.4209x-0.0051R=0.99785.
4. measure and get 0.5mL need testing solution, by the method under standard curve preparation, measure absorbance in accordance with the law.Calculate, the total triterpene of three batch samples and triterpene saponin content are respectively 6.09%, 5.80%, 5.50%, and known total triterpene and triterpene saponin content are more than 5.50%.
Embodiment 4 Fomes Fomentarius compound extract extracorporeal anti-tumor effects
Trophophase cell A549, Hela, QGY, DU145, HCT116, Eca109, MDA-MB-435S, SGC-7901, Saos-2 and U2os take the logarithm, cell trypsinization, adjust cell concentration, then cell suspension inoculation in 96 well culture plates, every hole 190 μ L, approximately 5000 cells.37 DEG C, 5%CO
2, under saturated humidity condition, cultivate 24h, then every hole adds the medicinal liquid of 10 μ L variable concentrations, continues to cultivate 48h.Outwell culture medium, every hole adds 100 μ LPBS and washes version once, then every hole adds the MTT solution (with the culture medium preparation of serum-free) of 100 μ L0.5mg/mL, after continuing to cultivate 4h, abandon supernatant, then every hole adds 150 μ LDMSO, the 10min that vibrates gently, absorbance (A) is surveyed in about 1h left and right at 570nm wavelength place by microplate reader, calculate IC
50.The IC of Fomes Fomentarius compound extract to A549, Hela, QGY, DU145, HCT116, Eca109, MDA-MB-435S, SGC-7901, Saos-2 and U2os
50be respectively 0.542,0.365,0.252,0.521,0.326,0.593,0.870,0.562,0.396 and 0.627mg/ml.
Embodiment 5 Fomes Fomentarius compound extract acute toxicity testings
Mtd test: 60 of the healthy KM mices (male and female half and half) of getting body weight 18～22g, be divided into immediately 3 groups, every group 20 (male and female half and half), twice (morning and afternoon are respectively once for every group of medicine that gavages variable concentrations, interval > 6h), each 0.5mL, observes the toxic reaction of animal in 7 days or has or not death.Taking death as MAIN OUTCOME MEASURES, if any death,, just not occur that dead group is as the criterion, dosage is now the maximum tolerated dose of mice.Then calculate tolerance multiple by formula, taking tolerate people's consumption oral 100 times above as safety, computing formula is as follows:
As a result, the maximum of every mice tolerates 0.2g the most, and amounting to crude drug amount is 4.58g, and the maximal tolerance times of mice is 100 times of 327 times of >, and its safety is described.
Embodiment 6 Fomes Fomentarius compound extracts suppress mouse tumor growth
(1) S180 transplanted tumor test: get well-grown S180 ascites tumor KM kind Mus, under aseptic condition, get ascites, with normal saline dilution in 1: 8, slowly mix, be seeded in allogeneic mice right fore oxter subcutaneous, every 0.2mL, after inoculation 24h, be divided at random 5 groups by body weight, every group 10, be respectively mould agent group, low dose group (50mg/kg), middle dosage group (100mg/kg), high dose group (150mg/kg) and positive controls (CTX, 30mg/kg).Model group lumbar injection every day 0.2mL normal saline, the medicinal liquid of administration group lumbar injection every day 0.2mL variable concentrations, positive controls lumbar injection every day 0.2mL cyclophosphamide solution (CTX).Administration strips tumor piece after 8 days, weigh, and calculates tumour inhibiting rate.The average tumor of tumour inhibiting rate (%)=(the average tumor weight of the average tumor weight-administration of model group group)/model group heavy × 100%.Result is as shown in table 2.
The inhibitory action of table 2 Fomes Fomentarius compound extract to S180 tumor-bearing mice tumor
Note：
*P＜0.05and
**P＜0.01vs blank control
(2) Lewis lung cancer transplanted tumor model: get the Lewis lung cancer C57BL/6 mice that tumor growth is good, get tumor piece under aseptic condition, reject peplos and downright bad part, add normal saline (1: 4) homogenate.Be seeded in allogeneic mice right fore oxter subcutaneous, every 0.2mL, after inoculation inoculation 24h, be divided at random 5 groups by body weight, every group 10, be respectively model group, low dose group (50mg/kg), middle dosage group (100mg/kg), high dose group (150mg/kg) and positive controls (CTX, 30mg/kg).Model group lumbar injection every day 0.2mL normal saline, the medicinal liquid of administration group lumbar injection every day 0.2mL variable concentrations, positive controls lumbar injection every day 0.2mL cyclophosphamide solution (CTX).Administration strips tumor piece after 15 days, weigh, and calculates tumour inhibiting rate.The average tumor of tumour inhibiting rate (%)=(the average tumor weight of the average tumor weight-administration of model group group)/model group heavy × 100%.Result is as shown in table 3.
The inhibitory action of table 3 water hoof compound extract to Lewis lung cancer tumor-bearing mice tumor
Note：
*P＜0.05and
**P＜0.01vs blank control
Embodiment 7 Fomes Fomentarius compound extract inducing apoptosis of tumour cell
The trophophase cell A549 that takes the logarithm, is inoculated in the single cell suspension 1.9mL of digestion counting in 6 orifice plates every hole approximately 1.5 × 10
5individual cell.After cell attachment, add the extract of 100 μ L variable concentrations, matched group adds the extract dissolve medium (culture medium of serum-free) of equivalent, after cultivating 24h, collect supernatant, cell washes twice with the PBS of pre-cooling, and trypsinization beats rear collecting cell, and the PBS of pre-cooling washes twice, cell suspends with the PBS containing 2% calf serum, add PI dyestuff, act on 5min on ice, by flow cytometer detection apoptosis situation.Result as shown in Figure 1.
Embodiment 8 Fomes Fomentarius compound extract stimulating effect immunostimulant
The force methods such as animal inoculation, grouping and administration are with embodiment 5, and administration finishes, and each treated animal is plucked eyeball and got blood, separation of serum; Strip Thymus and spleen, weigh, calculate thymus (spleen) index.Thymus (spleen) index=thymus (spleen) weight (mg)/body weight (g).Serum TNF-a assay: use ELISA test kit, adopt double fastener heart enzyme linked immunosorbent detection method, λ=450nm, according to standard A value curve plotting, reads the TNF-a content of sample to be detected from standard curve.Result is as shown in table 4.
Table 4-1 Fomes Fomentarius compound extract to S180 tumor-bearing mice thymus index, spleen index and
The impact that peripheral blood T NF-α generates
The impact that table 4-2 Fomes Fomentarius compound extract generates Lewis lung cancer tumor-bearing mice thymus index, spleen index and peripheral blood T NF-α
Note：
*P＜0.05and
**P＜0.01 vs blank control
The profit that the present invention proposes has remarkable antitumor efficacy and the low Fomes Fomentarius compound extract of toxicity and preparation method thereof, those skilled in the art can be by using for reference herein, obviously can not depart from content of the present invention, spirit and scope, appropriate change raw material, technique etc. realize this product.Special needs to be pointed out is, all similar replacements and change, be all regarded as in spirit of the present invention, scope and content.
Claims (6)
1. Fomes Fomentarius compound extract, is characterized in that the content of triterpenes and triterpene saponin is not less than 5.5%.
2. the preparation method of Fomes Fomentarius compound extract described in claim 1, is characterized in that: comprises the following steps,
Step 1: Fomes Fomentarius compound recipe is pulverized;
Step 2: the Fomes Fomentarius compound recipe after pulverizing adds water, heating and refluxing extraction is after stand at low temperature, centrifugal, supernatant concentrating under reduced pressure.Afterwards, concentrated solution adds ethanol again, and making ethanol final concentration is 80%, after stand at low temperature, centrifugal, gets supernatant, concentrating under reduced pressure again, and concentrated solution vacuum drying, to obtain final product.
3. the purposes of the compound extract of Fomes Fomentarius described in claim 1, it is characterized by: ((1) this extract is in vitro to human lung adenocarcinoma cell (A549), human cervical carcinoma cell (Hela), human liver cancer cell (QGY), Human Prostate Cancer Cells (DU145), human colon cancer cell (HCT116), human esophagus cancer cell (Eca109), human breast cancer cell (MDA-MB-435S), gastric carcinoma cells (SGC-7901), human osteosarcoma cell cell (Saos-2), the tumor cell lines such as human osteosarcoma cell (U2Os) have remarkable lethal effect.(2) this extract can effectively be induced the apoptosis of tumor cells including A549.(3) this extract has significant inhibitory action to murine sarcoma S180 and Lewis lung cancer transplanted tumor model tumor in vivo.(4) this extract can effectively improve tumor-bearing mice immunologic function, can make the thymus index of tumor-bearing mice and peripheral blood TNF-a content improve.
4. Fomes Fomentarius compound recipe according to claim 2, is characterized in that: Fomes fomentarius: phellin uspini: the weight ratio of Ganoderma lucidum (Leyss. Ex Fr.) Karst. is 1～10: 1～9: 1.
5. the compound extract of Fomes Fomentarius described in claim 2 mixes with pharmaceutically acceptable carrier, can be made into compound preparation.
6. compound preparation described in claim 5 comprises oral liquid, injection, suspensoid, pill, tablet, capsule, powder, granule or drop pill.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210355671.0A CN104055803A (en) | 2012-09-19 | 2012-09-19 | Fomes fomentarius compound extract and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210355671.0A CN104055803A (en) | 2012-09-19 | 2012-09-19 | Fomes fomentarius compound extract and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104055803A true CN104055803A (en) | 2014-09-24 |
Family
ID=51543922
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210355671.0A Pending CN104055803A (en) | 2012-09-19 | 2012-09-19 | Fomes fomentarius compound extract and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104055803A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104547444A (en) * | 2015-01-12 | 2015-04-29 | 赵玉梅 | Traditional Chinese medicine for treating phlegm-damp stagnation type gastric cancer and preparation method of traditional Chinese medicine |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1651058A (en) * | 2004-12-10 | 2005-08-10 | 陈康林 | Chinese medicine for treating tumour |
| CN101011425A (en) * | 2007-01-31 | 2007-08-08 | 陈康林 | Traditional Chinese medicine for treating esophagus cancer, gastric cancer, uterocarcinoma and hysteromyoma |
| CN101028296A (en) * | 2006-11-28 | 2007-09-05 | 陈康林 | Chinese medicine for treating tumor |
| CN102451198A (en) * | 2010-11-01 | 2012-05-16 | 刘峰 | Active traditional Chinese medicine formula for preventing cancers and preparation process thereof |
-
2012
- 2012-09-19 CN CN201210355671.0A patent/CN104055803A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1651058A (en) * | 2004-12-10 | 2005-08-10 | 陈康林 | Chinese medicine for treating tumour |
| CN101028296A (en) * | 2006-11-28 | 2007-09-05 | 陈康林 | Chinese medicine for treating tumor |
| CN101011425A (en) * | 2007-01-31 | 2007-08-08 | 陈康林 | Traditional Chinese medicine for treating esophagus cancer, gastric cancer, uterocarcinoma and hysteromyoma |
| CN102451198A (en) * | 2010-11-01 | 2012-05-16 | 刘峰 | Active traditional Chinese medicine formula for preventing cancers and preparation process thereof |
Non-Patent Citations (3)
| Title |
|---|
| 杨建宇等: "野生灵芝抗癌方推荐", 《2007国际中医药肿瘤大会会刊》 * |
| 陆勇芹等: "木蹄层孔菌化学成分及不同提取物体外抗肿瘤活性研究", 《西北林学院学报》 * |
| 陈康林: "《肿瘤与肝脏疾病治疗探索 肿瘤治疗的革命》", 30 November 2008, 中国古籍出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104547444A (en) * | 2015-01-12 | 2015-04-29 | 赵玉梅 | Traditional Chinese medicine for treating phlegm-damp stagnation type gastric cancer and preparation method of traditional Chinese medicine |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wachtel-Galor et al. | Ganoderma lucidum (Lingzhi or Reishi): a medicinal mushroom | |
| GB2428974A (en) | Botanical drug or dietary supplement | |
| EA014234B1 (en) | Composition for the treatment of hepatitis c or dietary supplement for prevention or alleviation hepatitis c symptoms | |
| CN103768151B (en) | Flos Osmanthi Fragrantis phenethyl alcohol glycoside extract is preparing the application in antiaging agent or health product | |
| Hu et al. | Efficacy evaluation of a Chinese bitter tea (Ilex latifolia Thunb.) via analyses of its main components | |
| KR101062172B1 (en) | Anti-cancer health food containing mushroom fermented ginseng powder | |
| CN109864991A (en) | Cryptotanshinone is preparing the application in Ph+ acute lymphoblastic leukemia chemical therapy sensitivity-enhancing | |
| Wei et al. | The immunomodulatory effects of active ingredients from Nigella sativa in RAW264. 7 cells through NF-κB/MAPK signaling pathways | |
| CN104274498B (en) | A kind of high efficiency anti-tumor Ganodenna Lucidum P.E and preparation method thereof | |
| CN108472502A (en) | For treating liver and maintaining composition, method and the pharmaceutical composition of liver health | |
| CN106389788B (en) | It is a kind of for enhancing the drug extract, preparation and preparation method of immunologic function of livestock and birds and anti-virus ability | |
| WO2015192758A1 (en) | Anti-tumor pharmaceutical application of pentacyclic triterpene saponin compounds of szechuan melandium root | |
| KR20110125295A (en) | Health assistance food used the euphorbia humifusa wild by main raw material and the manufacturing method therof | |
| Joshi | Antrodia camphorata with potential anti-cancerous activities: A review | |
| CN107519327A (en) | A kind of Phellinus Chinese medicine composition and its extracting method and the application in antineoplastic is prepared | |
| CN104055803A (en) | Fomes fomentarius compound extract and preparation method and application thereof | |
| TWI379685B (en) | ||
| Jones | Reishi mushroom: Ancient medicine in modern times | |
| KR101413283B1 (en) | The method for manufacturing panax ginseng composite by using the complex extract for the enhancement of immunity, and the panax ginseng composite made by the method | |
| CN102526283A (en) | Health-care medicine formula with functions of enhancing immunity, alleviating physical fatigue and resisting tumor | |
| CN106110305B (en) | Include pharmaceutical preparation and its application of cimicifugae foetidae saponin(e | |
| KR20160086457A (en) | Composition comprising alcohol extract of GamiBangkeehwangkeetang for preventing and treating a rheumatoid arthritis | |
| Abuarab et al. | Immunomodulatory and anticancer activities of barley bran grown in Jordan: An in vitro and in vivo study | |
| CN105597083B (en) | A kind of pair of liver kidney has health food of auxiliary protection function and preparation method thereof | |
| CN104415132A (en) | Compound formula for preparing soft capsule by pro-liposomes of ganoderma spore and freshwater mussel and preparation method thereof |
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: 20140924 |
|
| WD01 | Invention patent application deemed withdrawn after publication |