CN104248647B - A kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract and preparing the application in medicines resistant to liver cancer - Google Patents
A kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract and preparing the application in medicines resistant to liver cancer Download PDFInfo
- Publication number
- CN104248647B CN104248647B CN201310257032.5A CN201310257032A CN104248647B CN 104248647 B CN104248647 B CN 104248647B CN 201310257032 A CN201310257032 A CN 201310257032A CN 104248647 B CN104248647 B CN 104248647B
- Authority
- CN
- China
- Prior art keywords
- htj
- pers
- bull
- hericium erinaceus
- weight
- 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.)
- Active
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract and prepare the application in medicines resistant to liver cancer.This extract is that last vacuum lyophilization prepares by Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium concentrating under reduced pressure again after twice room temperature homogenate extraction.This extract has the effect of anti-hepatocarcinoma, can be used for preparing medicines resistant to liver cancer.
Description
Technical field
The invention belongs to biological pharmacy technical field, be specifically related to Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract and preparing the application in medicines resistant to liver cancer.
Background technology
Gastrointestinal cancer such as hepatocarcinoma, gastric cancer, knot, rectal cancer are one of modal cancers, estimate to account for 25% of all cancers according to ACS.Gastric cancer in the world sickness rate is positioned at the 4th in cancer, in state-owned very high incidence rate and mortality rate (death of 50% occurs in China), the treatment of gastrointestinal cancer comprises operation, chemotherapy, radiotherapy and immunotherapy.Cannot thoroughly treat although chemotherapy is the most frequently used method.In the decades in past, molecular biological development makes some progress in the treatment of gastric cancer, but drug resistance and toxicity still limit the progress of Therapeutic Method.Therefore, the new drug finding and develop the new anti-gastric cancer of high-efficiency low-toxicity is badly in need of.
Medicinal fungi refers to energy disease therapy, there is a class fungus of medical value, namely the such as multiclass material such as aminoacid, protein, vitamin, polysaccharide and glycoprotein, glycoside, alkaloid, sterols, Anthraquinones, flavonoid can be produced in mycelium, sporophore, sclerotium or spore, there is health-care effect to human body, disease is had to the fungus of prevention suppression or therapeutical effect.Fungus is as the medicinal history having more than 2000 year in China, the fungi-medicine such as Poria, Polyporus is just described in Shennong's Herbal, also the biological medicinal fungus of kind more than 20 has been recorded in Compendium of Material Medica, as medicinal and test, current China tradition confirms that the medicative fungus of tool is more than 400 kinds, mainly concentrates on load bacterium subphylum and Ascomycotina.But daily being widely used in clinical but only has more than 20 to plant, " Chinese Pharmacopoeia " version in 2010 has clearly included 7 kinds, and this explanation is still far from perfect to the research of biological medicinal fungus, and development space is larger.Existing research mainly concentrates on a few kind such as Ganoderma, Cordyceps, and mainly for fungus polysaccharide as the research of dosage form and Application and Development.
Along with the development of bioscience and technology, fungus, Acarasiales, lichens etc. can have been separated from plant kingdom, set up fungus circle separately.Except the Ganoderma had long enjoyed a good reputation at present, Poria, Cordyceps, Auricularia Sophorae kind, the kind that this boundary is selected in Chinese medicine is also less, but wherein higher fungus has thousands of kinds, will there will be this type of fungus medicines more from now on undoubtedly, and have a high potential, will greatly enrich Chinese medicine treasure-house.In addition, the fungus medicine based on medicinal fungi, except tradition application sporophore, the modern times have developed into production technologies such as adopting liquid (deep layer) fermentation (application fungus ball and fermentation liquid), solid fermentation (application mycoplasma).The mycelium of its main medicinal part of common mycoplasma that solid fermentation produces still just contained by it and secondary substance thereof, but it should be noted that medicinal fungi fruiting body is except self can be used as medicine, also in order to carry out Chinese traditional medicine biology engineering research, new drug can be developed into.And fermentation engineering, the especially distinctive solid fermentation engineering of China, the time that it exists existing more than 2000 year history, by long solid fermentation, creating biological metabolic product comprises biological enzyme material, and its medicinal mycoplasma generated is than liquid fermentation good drug efficacy.But solid fermentation existing defects, easy living contaminants, existing solid-fermented technique research is studies in China focus, the direction of main research is aseptic culture, second incubation, miscellaneous bacteria suppress, Chinese crude drug implements two-way solid-fermented technique as Nutrient medium, break through the bottleneck of solid-fermented technique, realize the extensive industrialization of biological medicinal fungus.
Hericium erinaceus (Bull. Ex Fr.) Pers. is a kind of hedgehog hydnum Cordycepps mushroom (fungus), the history of gastroenteropathy more than 2000 is used for the treatment of at Chinese medicine, a series of micromolecular compound is as erinacines, be considered to have neuranagenesis, and damaged nerve tissue can be repaired through blood brain barrier, Hericium erinaceus (Bull. Ex Fr.) Pers. also has antiulcer, antiinflammatory, antimicrobial, immunomodulating, improve liver function, defying age, reduces blood glucose and blood fat, improves anti-anoxia ability, increase heart export and improve the effects such as remarks circulation, the medical preparation comprising Hericium erinaceus (Bull. Ex Fr.) Pers. widely uses in China.
Active substance in Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium the most important thing is polysaccharide and glycoprotein, the domestic and international research to Hericium erinaceus polysaccharide at present shows, Hericium erinaceus (Bull. Ex Fr.) Pers. Polysaccharides has multiple biological activity and pharmacological action, the phagocytic function of macrophage can be strengthened, promote the formation of hemolysin, anti-leukopenia, blood sugar lowering, anticoagulation, antithrombotic, mutation and anti-ageingly to wait for a long time.Therefore, Hericium erinaceus (Bull. Ex Fr.) Pers. Polysaccharides enjoys people to pay close attention to, and becomes the focus of the area research such as molecular biology, medicine, Food Science and Application and Development in recent years." Hunan Province's Chinese crude drug standard " version in 2009,122-123 page describes Hericium erinaceus culture, and Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium of the present invention is the Hericium erinaceus culture that this standard is recorded.
Current, carry out screening and finding active component from higher fungus and culture thereof, be applied to antitumor, antiviral and treatment diabetes etc., not only becoming the developing direction of state key research new drug, is also the focus of world exploitation biological medicine.But, at present and have no the report mycelial for Hericium erinaceus (Bull. Ex Fr.) Pers. extract being applied to anti-hepatocarcinoma.
Summary of the invention
The present invention is intended to overcome the deficiencies in the prior art, a kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract is provided, this Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract of this Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract can kill people hepatocarcinoma HepG2 and Huh-7 cell in vitro and can suppress the growth of HepG2 and Huh-7 transplanted human hepatocellular carcinoma in vivo, therefore can be applicable to prepare resistive connection liver-cancer medicine.
The preparation method of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract of the present invention comprises the steps:
(1) get Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium, carry out first time water room temperature homogenate extraction, extract 15 ~ 25 minutes, then carry out second time water room temperature homogenate extraction, extract 10 ~ 20 minutes, for the first time in water room temperature homogenate extraction, the weight of water is 4 ~ 6 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight, is preferably 5 times; In second time water room temperature homogenate extraction, the weight of water is 2 ~ 4 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight, is preferably 3 times;
(2) supernatant obtained after extracting solution is centrifugal has been evaporated to Precipitation under 55 ~ 60 DEG C of conditions; By concentrated solution vacuum lyophilization under-60 ~-55 DEG C of conditions, both obtained Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract.
Wherein, the temperature of described water room temperature homogenate extraction is 20 ~ 50 DEG C.Described in step (2), extracting solution centrifugal condition is: rotating speed 3500 ~ 4500 revs/min, centrifugal 8 ~ 15 minutes.
Preferably, the preparation process of described extract also comprises: by the concentrated solution of step (2) again with weight be this concentrated solution weight 4 ~ 6 times and concentration of volume percent is the dissolve with ethanol of 70 ~ 80%, dissolving part is evaporated to without alcohol taste at 55 ~ 60 DEG C, by gained concentrate-60 ~-55 DEG C of vacuum lyophilizations.
Preferably, the preparation process of described extract also comprises: by the concentrated solution of step (2) again with weight be this concentrated solution weight 4 ~ 6 times and concentration of volume percent is the dissolve with ethanol of 70 ~ 80%, will not dissolve part-60 ~-55 DEG C of vacuum lyophilizations.
Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium of the present invention is " Hunan Province's Chinese crude drug standard " version in 2009, the Hericium erinaceus culture that 122-123 page is recorded, this Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium is hedgehog hydnum Cordycepps fungus Hericium erinaceus (Bull. Ex Fr.) Pers. Hericiumerinaceum(Bull.exFr.) mycelium of the Pers. drying composite of solid medium of growing nonparasitically upon another plant with it.
Room temperature homogenate extraction Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium of the present invention, under room temperature or room temperature (20-50 DEG C), does not heat, add hydrodynamic force making beating formula water-soluble go out effective ingredient, without temperature damage, can effective ingredient be dissolved but not destroy effective ingredient; 60 DEG C of concentrating under reduced pressure, can not destroy effective ingredient, particularly active enzyme material; In order to not affect regarding material, present invention employs vacuum lyophilization means.
The Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract of the present invention to gained has carried out the experiment suppressing people hepatocarcinoma HepG2 and Huh-7 cell and the anti-transplanted human hepatocellular carcinoma of animal, from Fig. 1 to Figure 22 and table 1 to the result of table 5, this Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract has the effect of anti-hepatocarcinoma, can be used for preparing medicines resistant to liver cancer.
Accompanying drawing explanation
Fig. 1: HTJ#2 inhibitory action to human hepatoma HepG2 cell; Concentration is 0.3125 – 20 mg/ml, 72 Hours drug process, and Thiazolyl blue (MTT) method measures cytoactive;
Fig. 2: HTJ#2A inhibitory action to human hepatoma HepG2 cell; Concentration is 0.3125 – 20 mg/ml, 72 Hours drug process, and Thiazolyl blue (MTT) method measures cytoactive;
Fig. 3: HTJ#5 inhibitory action to human hepatoma HepG2 cell; Concentration is 0.3125 – 20 mg/ml, 72 Hours drug process, and Thiazolyl blue (MTT) method measures cytoactive;
Fig. 4: HTJ#5A inhibitory action to human hepatoma HepG2 cell; Concentration is 0.3125 – 20 mg/ml, 72 Hours drug process, and Thiazolyl blue (MTT) method measures cytoactive;
Fig. 5: HTJ#2 inhibitory action to people hepatocarcinoma Huh-7 cell; Concentration is 0.3125 – 20 mg/ml, 72 Hours drug process, and Thiazolyl blue (MTT) method measures cytoactive;
Fig. 6: HTJ#2A inhibitory action to people hepatocarcinoma Huh-7 cell; Concentration is 0.3125 – 20 mg/ml, 72 Hours drug process, and Thiazolyl blue (MTT) method measures cytoactive;
Fig. 7: HTJ#5 inhibitory action to people hepatocarcinoma Huh-7 cell; Concentration is 0.3125 – 20 mg/ml, 72 Hours drug process, and Thiazolyl blue (MTT) method measures cytoactive;
Fig. 8: HTJ#5A inhibitory action to people hepatocarcinoma Huh-7 cell; Concentration is 0.3125 – 20 mg/ml, 72 Hours drug process, and Thiazolyl blue (MTT) method measures cytoactive;
Fig. 9: Fig. 9 A is the interior efficacy analysis to people hepatocarcinoma HepG2 transplanted tumor of HTJ#2 and 5-Fluorouracil body, and Fig. 9 B is the interior animal toxicity analysis to people hepatocarcinoma HepG2 transplanted tumor of HTJ#2 and 5-Fluorouracil body; Matched group, 5-Fluorouracil (30 mgs/kg/day, 5 days lumbar injections), HTJ#2 (200 mgs/kg/day, 5 days lumbar injections), with HTJ#2 (500 mgs/kg/day, 5 days oral), to Synergistic action and the toxicity of the SCID mice of lotus people hepatocarcinoma HepG2 transplanted tumor.Often organize 3 animals;
Figure 10: HTJ#2 and 5-Fluorouracil to the individual curative effect of people hepatocarcinoma HepG2 transplanted tumor; Matched group, 5-Fluorouracil (30 mgs/kg/day, lumbar injection), HTJ#2 (200 mgs/kg/day, lumbar injection), and HTJ#2 (500 mgs/kg/day, oral), by administration in continuous 5 days; Wherein, Figure 10 A is matched group, and Figure 10 B is 5-Fluorouracil (30 mgs/kg/day, lumbar injection), Figure 10 C is, HTJ#2 (200 mgs/kg/day, lumbar injection), Figure 10 D is HTJ#2 (500 mgs/kg/day, oral); In figure abscissa be the time (my god), vertical coordinate is tumor weight (milligram);
Figure 11: Figure 11 A is the interior efficacy analysis to people hepatocarcinoma HepG2 transplanted tumor of HTJ#2A and 5-Fluorouracil body, and Figure 11 B is the interior animal toxicity analysis to people hepatocarcinoma HepG2 transplanted tumor of HTJ#2A and 5-Fluorouracil body; Matched group, 5-Fluorouracil (30 mgs/kg/day, 5 days lumbar injections), HTJ#2A (200 mgs/kg/day, 5 days lumbar injections), with HTJ#2A (500 mgs/kg/day, 5 days oral), to Synergistic action and the toxicity of the SCID mice of lotus people hepatocarcinoma HepG2 transplanted tumor.Often organize 3-4 animal;
Figure 12: HTJ#2A and 5-Fluorouracil to the individual curative effect of people hepatocarcinoma HepG2 transplanted tumor; Matched group, 5-Fluorouracil (30 mgs/kg/day, lumbar injection), HTJ#2A (200 mgs/kg/day, lumbar injection), and HTJ#2A (500 mgs/kg/day, oral), by administration in continuous 5 days; Wherein, Figure 12 A is matched group, and Figure 12 B is 5-Fluorouracil (30 mgs/kg/day, lumbar injection), Figure 12 C is HTJ#2A (200 mgs/kg/day, lumbar injection), Figure 12 D is HTJ#2A (500 mgs/kg/day, oral); In figure abscissa be the time (my god), vertical coordinate is tumor weight (milligram);
Figure 13: Figure 13 A is the interior efficacy analysis to people hepatocarcinoma HepG2 transplanted tumor of HTJ#5A and 5-Fluorouracil body; Figure 13 B is the interior animal toxicity to people hepatocarcinoma HepG2 transplanted tumor of HTJ#5A and 5-Fluorouracil body; Matched group, 5-Fluorouracil (30 mgs/kg/day, 5 days lumbar injections), HTJ#5A (500 mgs/kg/day, 5 days lumbar injections), with HTJ#5A (500 mgs/kg/day, 5 days oral), to Synergistic action and the toxicity of the SCID mice of lotus people hepatocarcinoma HepG2 transplanted tumor.Often organize 3 animals;
Figure 14: HTJ#5A and 5-Fluorouracil to the individual curative effect of people hepatocarcinoma HepG2 transplanted tumor; Matched group, 5-Fluorouracil (30 mgs/kg/day, lumbar injection), HTJ#5A (500 mgs/kg/day, lumbar injection), and HTJ#5A (500 mgs/kg/day, oral), by administration in continuous 5 days; Wherein, Figure 14 A is matched group, and Figure 14 B is 5-Fluorouracil (30 mgs/kg/day, lumbar injection), Figure 14 C is HTJ#5A (500 mgs/kg/day, lumbar injection), Figure 14 D is HTJ#5A (500 mgs/kg/day, oral); In figure abscissa be the time (my god), vertical coordinate is tumor weight (milligram);
Figure 15: Figure 15 A is the interior efficacy analysis to people hepatocarcinoma HepG2 transplanted tumor of HTJ#5, HTJ#5A and 5-Fluorouracil body, and Figure 15 B is the interior animal toxicity to people hepatocarcinoma HepG2 transplanted tumor of HTJ#5, HTJ#5A and 5-Fluorouracil body; Matched group, 5-Fluorouracil (30 mgs/kg/day, 5 days lumbar injections), HTJ#5 (500 mgs/kg/day, 5 days oral), with HTJ#5A (500 mgs/kg/day, 5 days oral), to Synergistic action and the toxicity of the SCID mice of lotus people hepatocarcinoma HepG2 transplanted tumor.Often organize 5 animals;
Figure 16: HTJ#5, HTJ#5A and 5-Fluorouracil are to the individual curative effect of people hepatocarcinoma HepG2 transplanted tumor; Matched group, 5-Fluorouracil (30 mgs/kg/day, lumbar injection), HTJ#5 (500 mgs/kg/day, oral), and HTJ#5A (500 mgs/kg/day, oral), by administration in continuous 5 days; Wherein, Figure 16 A is matched group, and Figure 16 B is 5-Fluorouracil (30 mgs/kg/day, lumbar injection), Figure 16 C is HTJ#5 (500 mgs/kg/day, oral), Figure 16 D is HTJ#5A (500 mgs/kg/day, oral); In figure abscissa be the time (my god), vertical coordinate is tumor weight (milligram);
Figure 17: Figure 17 A is the interior efficacy analysis to people hepatocarcinoma HepG2 transplanted tumor of HTJ#5, HTJ#5A and 5-Fluorouracil body, and Figure 17 B is the interior animal toxicity analysis to people hepatocarcinoma HepG2 transplanted tumor of HTJ#5, HTJ#5A and 5-Fluorouracil body; Matched group, 5-Fluorouracil (25 mgs/kg/day, 5 days lumbar injections), HTJ#5 (1000 mgs/kg/day, 5 days oral), with HTJ#5A (1000 mgs/kg/day, 5 days oral), to Synergistic action and the toxicity of the SCID mice of lotus people hepatocarcinoma HepG2 transplanted tumor.Often organize 5 animals;
Figure 18: HTJ#5, HTJ#5A and 5-Fluorouracil are to the individual curative effect of people hepatocarcinoma HepG2 transplanted tumor; Matched group, 5-Fluorouracil (25 mgs/kg/day, lumbar injection), HTJ#5 (1000 mgs/kg/day, oral), and HTJ#5A (1000 mgs/kg/day, oral), by administration in continuous 5 days; Wherein, Figure 18 A is matched group, and Figure 18 B is 5-Fluorouracil (25 mgs/kg/day, lumbar injection), Figure 18 C is HTJ#5 (1000 mgs/kg/day, oral), Figure 18 DHTJ#5A (1000 mgs/kg/day, oral); In figure abscissa be the time (my god), vertical coordinate is tumor weight (milligram);
Figure 19: Figure 19 A is the interior efficacy analysis to people hepatocarcinoma Huh-7 transplanted tumor of HTJ#5, HTJ#5A and 5-Fluorouracil body, and Figure 19 B is the interior animal toxicity analysis to people hepatocarcinoma Huh-7 transplanted tumor of HTJ#5, HTJ#5A and 5-Fluorouracil body; Matched group, 5-Fluorouracil (25 mgs/kg/day, 5 days lumbar injections), HTJ#5 (1000 mgs/kg/day, 5 days oral), with HTJ#5A (1000 mgs/kg/day, 5 days oral), to Synergistic action and the toxicity of the SCID mice of lotus people hepatocarcinoma Huh-7 transplanted tumor.Often organize 5 animals;
Figure 20: HTJ#5, HTJ#5A and 5-Fluorouracil are to the individual curative effect of people hepatocarcinoma Huh-7 transplanted tumor; Matched group, 5-Fluorouracil (25 mgs/kg/day, lumbar injection), HTJ#5 (1000 mgs/kg/day, oral), and HTJ#5A (1000 mgs/kg/day, oral), by administration in continuous 5 days; Wherein, Figure 20 A is matched group, and Figure 20 B is 5-Fluorouracil (25 mgs/kg/day, lumbar injection), Figure 20 C is HTJ#5 (1000 mgs/kg/day, oral), Figure 20 DHTJ#5A (1000 mgs/kg/day, oral); In figure abscissa be the time (my god), vertical coordinate is tumor weight (milligram);
Figure 21: Figure 21 A is the interior efficacy analysis to people hepatocarcinoma Huh-7 transplanted tumor of HTJ#5, HTJ#5A and 5-Fluorouracil body, and Figure 21 B is the interior animal toxicity analysis to people hepatocarcinoma Huh-7 transplanted tumor of HTJ#5, HTJ#5A and 5-Fluorouracil body; Matched group, 5-Fluorouracil (25 mgs/kg/day, 5 days lumbar injections), HTJ#5 (1000 mgs/kg/day, 5 days oral), with HTJ#5A (1000 mgs/kg/day, 5 days oral), to Synergistic action and the toxicity of the SCID mice of lotus people hepatocarcinoma Huh-7 transplanted tumor.Often organize 5 animals;
Figure 22: HTJ#5, HTJ#5A and 5-Fluorouracil are to the individual curative effect of people hepatocarcinoma Huh-7 transplanted tumor; Matched group, 5-Fluorouracil (25 mgs/kg/day, lumbar injection), HTJ#5 (1000 mgs/kg/day, oral), and HTJ#5A (1000 mgs/kg/day, oral), by administration in continuous 5 days; Wherein, Figure 22 A is matched group, and Figure 22 B is 5-Fluorouracil (25 mgs/kg/day, lumbar injection), Figure 22 C is HTJ#5 (1000 mgs/kg/day, oral), Figure 22 DHTJ#5A (1000 mgs/kg/day, oral); In figure abscissa be the time (my god), vertical coordinate is tumor weight (milligram).
Detailed description of the invention
Below in conjunction with drawings and Examples, the present invention is further illustrated.
Embodiment 1
A kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract, its preparation method comprises the steps:
(1) get Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium, carry out first time water room temperature homogenate extraction, extract 20 minutes, then carry out second time water room temperature homogenate extraction, extract 15 minutes; For the first time in water room temperature homogenate extraction, the weight of water is 5 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight; In second time water room temperature homogenate extraction, the weight of water is 3 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight;
(2) supernatant obtained after centrifugal for the extracting solution obtained after above-mentioned extracted twice is evaporated to Precipitation under 60 DEG C of conditions and obtains concentrated solution; By concentrated solution vacuum lyophilization under-60 DEG C of conditions, obtain Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract.
Wherein, the temperature of described water room temperature homogenate extraction is 30 DEG C.Described in step (2), extracting solution centrifugal condition is: rotating speed 4000 revs/min, centrifugal 10 minutes.
Embodiment 2
A kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract, its preparation method comprises the steps:
(1) get Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium, carry out first time water room temperature homogenate extraction, extract 20 minutes, then carry out second time water room temperature homogenate extraction, extract 15 minutes; For the first time in water room temperature homogenate extraction, the weight of water is 5 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight; In second time water room temperature homogenate extraction, the weight of water is 3 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight;
(2) supernatant obtained after centrifugal for the extracting solution obtained after above-mentioned extracted twice is evaporated to Precipitation under 60 DEG C of conditions and obtains concentrated solution;
(3) by the concentrated solution of step (2) again with weight be this concentrated solution weight 5 times and concentration of volume percent is the dissolve with ethanol of 80%, dissolving part is evaporated to without alcohol taste at 60 DEG C, by gained concentrate-60 DEG C of vacuum lyophilizations, obtain the Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract extracted further.
Wherein, the temperature of described water room temperature homogenate extraction is 30 DEG C.Described in step (2), extracting solution centrifugal condition is: rotating speed 4000 revs/min, centrifugal 10 minutes.
Embodiment 3
A kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract, its preparation method comprises the steps:
(1) get Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium, carry out first time water room temperature homogenate extraction, extract 20 minutes, then carry out second time water room temperature homogenate extraction, extract 15 minutes; For the first time in water room temperature homogenate extraction, the weight of water is 5 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight; In second time water room temperature homogenate extraction, the weight of water is 3 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight;
(2) supernatant obtained after centrifugal for the extracting solution obtained after above-mentioned extracted twice is evaporated to Precipitation under 60 DEG C of conditions and obtains concentrated solution;
(3) by the concentrated solution of step (2) again with weight be this concentrated solution weight 5 times and concentration of volume percent is the dissolve with ethanol of 80%, by insoluble part-60 DEG C of vacuum lyophilizations, obtain the Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract extracted further.
Wherein, the temperature of described water room temperature homogenate extraction is 30 DEG C.Described in step (2), extracting solution centrifugal condition is: rotating speed 4000 revs/min, centrifugal 10 minutes.
Embodiment 4
The inhibitory action of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract Functional Validation Test-to people hepatocarcinoma HepG2 and Huh-7 cell
One, materials and methods
1, cell and cultivation: people hepatocarcinoma HepG2 and Huh-7 cell are from American Type Culture institute (AmericanTypeCultureCollection, ATCC; Manassas, VA, UAS).HepG2 and Huh-7 cell is raised in DMEM culture fluid, containing 10% calf serum (AtlantaBiological, Lawrenceville, GA, USA), the penicillin of 100 units per ml, with the streptomycin (Invitrogen of 0.1 mcg/ml, GrandIsland, NY, USA).Cell is in 37 DEG C, 5%CO
2cultivate in incubator.
2, medicine: the Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract finally obtained by embodiment 1 preparation method is divided into two groups, called after HTJ#2 and HTJ#2A respectively, the Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract called after HTJ#5 finally obtained by embodiment 2 preparation method, the Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract called after HTJ#5A finally obtained by embodiment 3 preparation method, is dissolved in the DMEM culture fluid containing 0.1%DMSO respectively by aforementioned Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract.
3, drug level and scheme: HTJ#2, HTJ#2A, HTJ#5, and HTJ#5A drug level is in 0.3125 – 20 mg/ml, continuous 72 Hours drug process.
4. suppress people hepatocarcinoma HepG2 and Huh-7 cells growth activity: 72 Hours drug process, then cell uses Thiazolyl blue (MTT) method to measure cytoactive.
Two, experimental result
1.HTJ#2, HTJ#2A, HTJ#5 and HTJ#5A are to the growth inhibited effect of human hepatoma HepG2 cell.
The human hepatoma HepG2 cell that first we use 13 kinds of different HTJs (#2, #2A, #2B, #4, #5, #5A, #5B, #6, #7, #8, #9, #9-1, and #9-2) to process In vitro culture observes the growth in vitro inhibitory action to hepatoma carcinoma cell after 72 hours.Experimental result shows that all HTJs effectively can suppress the growth of human hepatoma HepG2 cell, its inhibitory action amount effect curve (data do not show).The wherein inhibitory action of HTJ#2, HTJ#2A, HTJ#5 and HTJ#5A comparatively strong (Fig. 1-4 and table 1), so we are used for interior animal experiment.Fig. 1 and Fig. 2 data show that HTJ#2 and HTJ#2A is very strong to the inhibitory action of human hepatoma HepG2 cell, its half casualty-producing concentrations (IC
50) be respectively 0.40 ± 0.04 and 0.35 ± 0.02 mg/ml (table 1), the HepG2 cell being greater than 80% can be killed and wounded in 0.625 mg/ml.The inhibitory action of HTJ#5 and HTJ#5A to HepG2 cell is weak compared with HTJ#2 and HTJ#2A, to the half casualty-producing concentrations (IC of HepG2 cell
50) being respectively 2.50 ± 0.25 and 2.00 ± 0.25 mg/ml (table 1), HTJ#5 can kill and wound the HepG2 cell being greater than 70% in 5.0 mg/ml, HTJ#5A can kill and wound the HepG2 cell (Fig. 3 and Fig. 4) being greater than 85% in 5.0 mg/ml.
2.HTJ#2HTJ#2A, HTJ#5 and HTJ#5A are to the growth inhibited effect of people hepatocarcinoma Huh-7 cell.
Equally, we also using 13 kinds of different HTJs (#2, #2A, #2B, #4, #5, #5A, #5B, #6, #7, #8, #9, #9-1, and #9-2) process the another kind of human liver cancer cell Huh-7 of In vitro culture, observe the growth in vitro inhibitory action to hepatoma carcinoma cell after 72 hours.Experimental result shows that all HTJs also effectively can suppress the growth of people hepatocarcinoma Huh-7 cell, its inhibitory action amount effect curve (data do not show).Fig. 5 to Fig. 8 and table 1 shows HTJ#2, and HTJ#2A, HTJ#5 and HTJ#5A are to the inhibitory action of Huh-7 cell.Fig. 1 and Fig. 2 data show that HTJ#2 and HTJ#2A is also very strong to the inhibitory action of people hepatocarcinoma Huh-7 cell, its half casualty-producing concentrations (IC
50) being respectively 0.50 ± 0.03 and 0.20 ± 0.01 mg/ml (table 1), HTJ#2 can kill and wound the Huh-7 cell being greater than 90% in 1.25 mg/ml.HTJ#2A can kill and wound the HepG2 cell being greater than 80% in 0.625 mg/ml.Equally, the inhibitory action of HTJ#5 and HTJ#5A to Huh-7 cell is weak compared with HTJ#2 and HTJ#2A, but the inhibitory action of comparison HepG2 cell is strong, to the half casualty-producing concentrations (IC of Huh-7 cell
50) being respectively 0.80 ± 0.08 and 1.5 ± 0.28 mg/ml (table 1), HTJ#5 can kill and wound the Huh-7 cell being greater than 70% in 1.25 mg/ml, HTJ#5A can kill and wound the Huh-7 cell (Fig. 7 and Fig. 8) being greater than 70% in 5.0 mg/ml.
Embodiment 5
Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract Functional Validation Test
One, materials and methods
1. animal: the female Mus of SCID (weight 18 ~ 22g) of 8 ~ 12 weeks, from RoswellParkCancerInstitute (Buffalo, NY) and by regulation raising.
2. medicine: the Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract finally obtained by embodiment 1 or 2 preparation method, called after HTJ#2, HTJ#2A, HTJ#5 and HTJ#5A is dissolved in the physiological saline solution containing 10%DMSO, 5-Fluorouracil (5-FU:50mg/ml) is purchased from Hoffmann-LaRoche, Inc (Nutley, NJ) dilutes with physiological saline solution.
3. tumor cell: HepG2 and Huh-7 hepatoma carcinoma cell.First the foundation of transplanted tumor injected about 1000000 cultured cells and transplanted through number generation.
4. drug dose and scheme: HTJ#2 is oral or inject 200-500 mg/kg/day, HTJ#2A is oral or inject 200-500 mg/kg/day, HTJ#5 is oral or inject 500-1000 mg/kg/day, HTJ#5A oral 500-1000 mg/kg/day, 5-Fluorouracil 25-30 mg/kg/day, intraperitoneal injection, once a day, 5 days courses for the treatment of.Matched group is oral or inject 10%DMSO solution, and it is identical with experimental group that mice every 20g body weight gives 200-400 μ l().Often organize 3-5 mice, in part Experiment, the transplanted tumor that same Mus lotus two kinds is different.
5. with the axis of vernier caliper measurement tumor, (L represents major axis, W represents minor axis) tumor weight is calculated as follows: tumor weight=1/2 (LxW2) (mm). (1mg=1mm3), the every day same time carries out measurement of tumor, 3-4 time weekly.
6. maximum tolerated dose (MTD) and toxicity assessment: maximum tolerated dose is defined as administration and causes Mouse Weight to alleviate about 20%, and medicine can not be caused to cause the dosage of dead mouse.Accepting front ten days after Drug therapy, measure tumor and Mouse Weight every day, is then measure once for every two days.
7. anti-tumor activity: antitumor activity evaluation index, maximum Tumor growth inhibition, calculates with the exemplary embodiment lock of the average tumor weight for the treatment of group and matched group; Tumor doubling time, the average time needed for the twice that tumor weight rises to initial weight; Tumor section reaction (PR), when tumor size reduces half (to initial 50% or more); Tumor complete reaction (CR), when detecting tumor (cases of complete remission); Cure after being defined as Drug therapy and at least 60 days, can't detect tumor (reach tumor complete reaction to keep at least 60 days, now general tumor can not recur, and animal no longer retains).
Two, experimental result
1.HTJ#2, HTJ#2A, HTJ#5, HTJ#5A and 5-Fluorouracil are to the anti-tumor activity of the SCID mice of lotus HepG2 people transplanted human hepatocellular carcinoma and animal toxicity
First we have studied HTJ#2, and HTJ#2A, HTJ#5A and 5-Fluorouracil are to the anti-tumor effect of the SCID mice of lotus people transplanted human hepatocellular carcinoma and animal toxicity.The test data display Vehicle controls group of Fig. 9-14 and table 2,5-Fluorouracil dosage is 30 mgs/kg/day (lumbar injections), HTJ#2 and HTJ#2A dosage is 200 mgs/kg/day (lumbar injections), with 500 mgs/kg/day (oral), HTJ#5A dosage is 500 mgs/kg/day (lumbar injections or oral), once a day, the anti-tumor activity of the SCID mice to lotus HepG2 hepatocarcinoma of continuous 5 days and toxicity.The use of dosage has resisted straight colon cancer and anti-incidence cancer test dose before.Result of the test shows that HepG2 Growth of Transplanted Hepatocarcinoma in Mice mean doubling time is 7.6 ± 1.0 days.Than the straight colon cancer of SW620 and FaDu incidence cancer growth of xenografted slow.5-Fluorouracil has gentle Tumor suppression active, and inhibition rate of tumor growth is 52.7 ± 6.7%, and tumour doubling time extends to 11.3 ± 0.7 days, and causes average mice body weight to decline 17%.Lumbar injection HTJ#2200 mg/kg/day, inhibition rate of tumor growth is 71.1 ± 2.4%, and tumour doubling time extends to 18.9 ± 0.7 days, causes average mice body weight to decline 10.3%.Oral 500 mgs/kg of/day HTJ#2, with lumbar injection 200 mgs/kg of/day HTJ#2, there is identical antitumor activity, inhibition rate of tumor growth is 68.6 ± 5.6%, tumour doubling time extends to 16.8 ± 1.4 days, causes average mice body weight decline 10.2% (figure .9-10 and table 2).The anti-HepG2 tumor activity of HTJ#2A is poorer than HTJ#2, and similar to 5-Fluorouracil, data are shown in Figure 11-12 and table 2.Result shows lumbar injection HTJ#2A200 mg/kg/day, and inhibition rate of tumor growth is 46.2 ± 9.8%, and tumour doubling time is 12.3 ± 0.6 days, causes average mice body weight to decline 12.4%.Oral HTJ#2A500 mg/kg/day, inhibition rate of tumor growth is 52.7 ± 2.1%, and tumour doubling time is 13.2 ± 0.8 days, causes average mice body weight decline 8.3% (Figure 11-12 and table 2).The Tumor suppression of HTJ#5A is active similar to HTJ#2, lumbar injection HTJ#5A500 mg/kg/day, and inhibition rate of tumor growth is 72.1 ± 2.0%, and tumour doubling time extends to 16.9 ± 0.4 days, and average mice body weight declines 5.7%.Oral 500 mgs/kg of/day HTJ#5A, with lumbar injection 200 mgs/kg of/day HTJ#2, there is identical antitumor activity, inhibition rate of tumor growth is 69.3 ± 11.1%, and tumour doubling time extends to 18.6 ± 1.7 days, average mice body weight decline 8.3% (figure .13-14 and table 2).
Then, we are repeated anti-hepatocarcinoma HepG2 transplanted tumor test-Vehicle controls group, 5-Fluorouracil dosage 30 mgs/kg/day (lumbar injection), to increase HTJ#5A500 mg/kg/day (oral) with HTJ#5500 mg/kg/day (oral), because test above knows HTJ#2, the antineoplastic effect of the oral and drug administration by injection approach of HTJ#2A with HTJ#5A is identical, so we have only carried out the test of HTJ#5 and HTJ#5A oral medication.HepG2 growth of xenografted is slower than test last time, and it is 7.6 ± 1.0 that tumour doubling time about 10.5 ± 0.9(tested last time) sky.The anti-tumor activity of 5-Fluorouracil is stronger than last time, and inhibition rate of tumor growth is 71.4 ± 2.2%, and tumour doubling time extends to 17.6 ± 4.0 days, and animal toxicity also all increases causes average mice body weight to decline 19.6%, and reaches the partial reaction (PR) of 60%.HTJ#5 oral dose 500 mgs/kg/day, tumor control rate is for about 70.0 ± 23.3% but tumour doubling time is longer than 5-Fluorouracil reaches about 21.0 ± 7.2 days, and there are the partial reaction (PR) of 40% and the complete reaction (CR of 20%, cure), average mice body weight declines and only has 9.7%.During HTJ#5A oral dose 500mg/kg/ days, tumor control rate is more by force that 77.6 ± 15.5% tumour doubling time reach 19.0 ± 6.1 days, and have the partial reaction (PR) of 80%, average mice body weight declines and only has 8.9%% (figure .15-16 and table 3).
We have also carried out the test of third time anti-hepatocarcinoma HepG2 transplanted tumor, the dosage of 5-Fluorouracil is reduced to 25 mgs/kg/day from 30 mgs/kg/day, and HTJ#5 and HTJ#5A dosage is increased 1000 mgs/kg/day from 500 mgs/kg/day, an increase anti-tumor activity can be entered to observe.Data are shown in Figure 17-18 and table 4.Data are visible, tumor growth is identical with primary test faster than secondary test, doubling time is 7.5 ± 0.4 days, the 5-Fluorouracil of 25 mgs/kg of dosage is lower than the anti-tumor activity of 30 mgs/kg of dosage, tumor control rate 49.0 ± 5.7%, tumour doubling time about 8.6 ± 0.4 days, does not obtain complete reaction (CR) or partial reaction (PR), toxicity also decreases simultaneously, and average mice body weight reduces by 15.1 ± 4.5%.HTJ#5 anti-tumor activity is better than 5-Fluorouracil activity, and tumor control rate is 66.8 ± 13.0%, the doubling time about 14.2 ± 1.8, and obtain the partial reaction (PR) of 20%, body weight is reduced to 8.2 ± 2.6%.The dosage (from 500 to 1000 mgs/kg/days) improving HTJ#5 does not improve anti-tumor activity and toxicity.The anti-tumor activity of HTJ#5A is better than HTJ#5, and tumor control rate reaches 70.4 ± 12.6%, and tumour doubling time is 16.3 ± 1.6 days, and obtain the partial reaction (PR) of 20%, average mice body weight is reduced to 6.4 ± 4.5%.Equally, better anti-tumor activity and toxicity (figure .17-18 and table 4) is had when being not 500 mgs/kg/day than dosage when HTJ#5A dosage is 1000 mgs/kg of/day.
2.HTJ#5, HTJ#5A and 5-Fluorouracil are to the anti-tumor activity of the SCID mice of lotus Huh-7 people transplanted human hepatocellular carcinoma and animal toxicity.
We have also investigated HTJ#5(1000 mg/kg/day), HTJ#5A(1000 mg/kg/day), and 5-Fluorouracil (25 mgs/kg/day) is to the effect of the SCID mice of another kind of transplanted human hepatocellular carcinoma Huh-7.Data are shown in Figure 19-20 and table 5.Data show that the Huh-7 transplanted tumor doubling time is 7.6 ± 0.3 days, and its speed of growth is similar to HepG2 transplanted tumor or slightly fast.Although 5-Fluorouracil is when 25 mgs/kg of/day dosage, initial anti-Huh-7 tumor activity obvious, growth inhibition ratio be 72.1 ± 3.9 (be 49% to HepG2), after stopped treatment, tumor ramp.Therefore do not observe obvious tumor growth delay phenomenon.Tumour doubling time is 7.8 ± 0.4 days, does not obtain complete reaction (CR) or partial reaction (PR), and simultaneously toxicity is also mg/kg/day low than 30, and average mice body weight reduces by 15.1 ± 4.5%.Oral HTJ#5 and HTJ#5A be ratio injection 5-Fluorouracil 25 mgs/kg/day 1000 mgs/kg of/day time, its anti-Huh-7 tumor promotion is stronger, growth inhibition ratio is respectively 80.4 ± 1.9 and 87.6 ± 5.0, and particularly the tumor growth doubling time reaches 14.2 ± 1.6 and 15.2 ± 1.5 days (P<0.01) respectively.And obtaining the partial reaction (PR) of 40% and 60% respectively, average mice body weight reduction is respectively 8.2 ± 2.6% and 6.4 ± 4.5%.Compare with HepG2 transplanted tumor, Huh-7 transplanted human hepatocellular carcinoma to 5-Fluorouracil, HTJ#5, and the primary response of HTJ#5A more obviously but curative effect be difficult to lastingly.
Then, we are repeated anti-hepatocarcinoma Huh-7 transplanted tumor test-Vehicle controls group, keep HTJ#5 and HTJ#5A dosage 1000 mgs/kg/day (oral), and increase 5-Fluorouracil dosage to 30 mg/kg/day (lumbar injection).Result of the test data are shown in Figure 21-22 and table 6.Result shows that Huh-7 growth of xenografted was tested faster than last time, and it is 7.6 ± 0.3 that tumour doubling time about 6.4 ± 0.3(tested last time) sky.5-Fluorouracil is active similar to 25 mgs/kg/day the anti-Huh-7 transplanted tumor of 30 mgs/kg/day, individual variation is large, inhibition rate of tumor growth is 67.6 ± 20.5%, tumour doubling time to be prolonged than 25 mg/kg/day long, to 8.9 ± 0.6 days, but do not obtain complete reaction (CR) or partial reaction (PR), animal toxicity is all increases also, average mice body weight declines 19.6 ± 2.7%.The anti-tumor curative effect of HTJ#5 with HTJ#5A was tested similar to last time, tumor control rate is respectively 79.3 ± 13.0% and 75.2 ± 6.2%, tumour doubling time is respectively 14.6 ± 0.9 and 14.2 ± 0.7 days, and obtain the partial reaction (PR) of 60% and 40% respectively, average mice body weight declines and only has 5.9 ± 5.2% and 5.5 ± 2.8% (figure .21-22 and tables 6).
Conclusion
1.HTJ#2, HTJ#2A, HTJ#5 and HTJ#5A have obvious inhibitory action to human hepatoma HepG2 cell in vitro.
2.HTJ#2, HTJ#2A, HTJ#5 and HTJ#5A have obvious inhibitory action to people hepatocarcinoma Huh-7 cell in vitro.
3..HTJ#2, HTJ#2A, HTJ#5, HTJ#5A and 5-Fluorouracil have certain anti-tumor curative effect, HTJ#2 to people's cancer liver HepG2 transplanted tumor in vivo, HTJ#5, stronger than the anti-tumor activity of 5-Fluorouracil with HTJ#5A, the anti-tumor activity of HTJ#2A is than HTJ#2, HTJ#5, poor with HTJ#5A, similar to the anti-tumor activity of 5-Fluorouracil.
4. 5-Fluorouracil is mg/kg/day better to the anti-tumor activity of cancer liver HepG2 transplanted tumor than 25 at 30 mgs/kg/day.
5. oral HTJ#2, HTJ#2A and HTJ#5A and lumbar injection HTJ#2, HTJ#2A and HTJ#A5 have identical anti-tumor activity to people's cancer liver HepG2 transplanted tumor.
6. oral 500 mgs/kg of/day HTJ#5 and HTJ#5A have identical anti-tumor activity with oral 1000 mgs/kg/day to cancer liver HepG2 transplanted tumor.
7., although 5-Fluorouracil is at 25-30 mg/kg/day, initial anti-Huh-7 tumor activity obvious, it is 70%, after stopped treatment that growth inhibition ratio is greater than, tumor ramp, therefore does not observe obvious tumor growth delay phenomenon.
The curative effect of the anti-Huh-7 tumour transplatation tumor of 8.HTJ#5 and HTJ#5A is better than 5-Fluorouracil, and tumor growth obviously postpones.
Table 1: sum up HTJ#2, HTJ#2A, HTJ#5 and HTJ#5A to the half casualty-producing concentrations (IC50) of HepG2 and Huh-7 human liver cancer cell
72 Hours drug process, Thiazolyl blue (MTT) method measures cytoactive.Often organize 3-4 sample.*Mean±SD.
Table 2: summary HTJ#2, HTJ#2A, HTJ#5A and 5-Fluorouracil are to the anti-tumor curative effect of HepG2 hepatocarcinoma and animal toxicity
Matched group every day by the every 20 grams of normal saline giving 0.2 milliliter of mouse body weight containing 10% DMSO.Each experimental group uses 5 mouse.Transplanted tumor starts treatment after transplanting and reaching about 190 to 220 milligrams in 7 days.
Table 3: summary HTJ#5, HTJ#5A and 5-Fluorouracil are to the anti-tumor curative effect of HepG2 hepatocarcinoma and animal toxicity
Matched group every day by the every 20 grams of normal saline giving 0.2 milliliter of mouse body weight containing 10% DMSO.Each experimental group uses 5 mouse.Transplanted tumor starts treatment after transplanting and reaching about 180 to 200 milligrams in 7 days.
Table 4: summary HTJ#5, HTJ#5A and 5-Fluorouracil are to the anti-tumor curative effect of HepG2 hepatocarcinoma and animal toxicity
Matched group every day by the every 20 grams of normal saline giving 0.2 milliliter of mouse body weight containing 10% DMSO.Each experimental group uses 5 mouse.Transplanted tumor starts treatment after transplanting and reaching about 180 to 200 milligrams in 7 days.
Table 5: summary HTJ#5, HTJ#5A and 5-Fluorouracil are to the anti-tumor curative effect of Huh7 hepatocarcinoma and animal toxicity
Matched group every day by the every 20 grams of normal saline giving 0.2 milliliter of mouse body weight containing 10% DMSO.Each experimental group uses 5 mouse.Transplanted tumor starts treatment after transplanting and reaching about 180 to 200 milligrams in 7 days.
Table 6: summary HTJ#5, HTJ#5A and 5-Fluorouracil are to the anti-tumor curative effect of Huh-7 hepatocarcinoma and animal toxicity
Matched group every day by the every 20 grams of normal saline giving 0.2 milliliter of mouse body weight containing 10% DMSO.Each experimental group uses 5 mouse.Transplanted tumor starts treatment after transplanting and reaching about 180 to 200 milligrams in 7 days.
Claims (2)
1. a Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract is preparing the application in medicines resistant to liver cancer; The preparation of described Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract comprises the steps:
(1) get Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium, carry out first time water room temperature homogenate extraction, extract 15 ~ 25 minutes, then carry out second time water room temperature homogenate extraction, extract 10 ~ 20 minutes; For the first time in water room temperature homogenate extraction, the weight of water is 4 ~ 6 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight; In second time water room temperature homogenate extraction, the weight of water is 2 ~ 4 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight; Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium is hedgehog hydnum Cordycepps fungus Hericium erinaceus (Bull. Ex Fr.) Pers. Hericiumerinaceum(Bull.exFr.) mycelium of the Pers. drying composite of solid medium of growing nonparasitically upon another plant with it; The temperature of described water room temperature homogenate extraction is 20 ~ 50 DEG C;
(2) supernatant obtained after centrifugal for the extracting solution obtained after above-mentioned extracted twice is evaporated to Precipitation under 55 ~ 60 DEG C of conditions and obtains concentrated solution; By concentrated solution again with weight be this concentrated solution weight 4 ~ 6 times and concentration of volume percent is the dissolve with ethanol of 70 ~ 80%, dissolving part is evaporated to without alcohol taste at 55 ~ 60 DEG C, by gained concentrate-60 ~-55 DEG C of vacuum lyophilizations, obtain Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract, or by concentrated solution again with weight be this concentrated solution weight 4 ~ 6 times and concentration of volume percent is the dissolve with ethanol of 70 ~ 80%, part will not be dissolved-60 ~-55 DEG C of vacuum lyophilizations, obtain Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract; Described extracting solution centrifugal condition is: rotating speed 3500 ~ 4500 revs/min, centrifugal 8 ~ 15 minutes.
2. apply as claimed in claim 1, it is characterized in that, in Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract preparation method in first time water room temperature homogenate extraction, the weight of water is 5 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight; In second time water room temperature homogenate extraction, the weight of water is 3 times of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium weight.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310257032.5A CN104248647B (en) | 2013-06-25 | 2013-06-25 | A kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract and preparing the application in medicines resistant to liver cancer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310257032.5A CN104248647B (en) | 2013-06-25 | 2013-06-25 | A kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract and preparing the application in medicines resistant to liver cancer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104248647A CN104248647A (en) | 2014-12-31 |
| CN104248647B true CN104248647B (en) | 2016-04-13 |
Family
ID=52184123
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310257032.5A Active CN104248647B (en) | 2013-06-25 | 2013-06-25 | A kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract and preparing the application in medicines resistant to liver cancer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104248647B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105495621A (en) * | 2015-12-17 | 2016-04-20 | 重庆希尔安药业有限公司 | Health-care food used for protecting liver and stomach and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1546533A (en) * | 2003-12-03 | 2004-11-17 | 高其品 | Hericium erinaceus sulfate proteoglycan, preparation method thereof and pharmaceutical compositions with the same as active component |
| CN1813819A (en) * | 2005-12-09 | 2006-08-09 | 南京老山药业股份有限公司 | Hedgehog hydnum fruiting body or hyphostroma and culture extract and formulation and preparing method |
-
2013
- 2013-06-25 CN CN201310257032.5A patent/CN104248647B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1546533A (en) * | 2003-12-03 | 2004-11-17 | 高其品 | Hericium erinaceus sulfate proteoglycan, preparation method thereof and pharmaceutical compositions with the same as active component |
| CN1813819A (en) * | 2005-12-09 | 2006-08-09 | 南京老山药业股份有限公司 | Hedgehog hydnum fruiting body or hyphostroma and culture extract and formulation and preparing method |
Non-Patent Citations (2)
| Title |
|---|
| 小刺猴头菌水溶性小分子提取物HPLC指纹图谱探讨;綦天华等;《中国生化药物杂志》;20120229;第33卷(第1期);第8-11页 * |
| 猴头菌小分子活性成分研究进展;尚晓冬等;《食用菌学报》;20120331;第19卷(第1期);第79-84页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104248647A (en) | 2014-12-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| TWI598104B (en) | Use of Antrodia cinnamomea extract to improve side effects of chemotherapy | |
| CN101019893A (en) | Process of enriching effective antitumor part of periplaneta americana with polyamide | |
| CN101773625B (en) | Anticancerogen, preparation method and application thereof | |
| CN104224855A (en) | Use of hericium erinaceus hyphae extract in preparation of drug for resisting stomach cancer | |
| CN104248647B (en) | A kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract and preparing the application in medicines resistant to liver cancer | |
| CN108570116B (en) | Pleurotus citrinopileatus polysaccharide, preparation method and medical application in preventing and treating diabetes | |
| CN104224856B (en) | Hericium erinaceus mycelium extract and application thereof in preparation of head-neck cancer resistant drugs | |
| CN104224857B (en) | A kind of Hericium erinaceus hypha extract is preparing the application in resisting straight colon cancer drug | |
| US11065293B2 (en) | Pharmaceutical composition for decreasing the side effects of cancer drug, and manufacturing method and uses thereof | |
| CN103230418B (en) | Mycelium extract for helicobacter pylori-resistant biological medicine and preparation technology thereof | |
| CN102078600B (en) | Anti-cancer compound ganoderma composition, application thereof and pharmaceutical composition containing same | |
| CN105535011A (en) | Applications of mushroom mycelium polysaccharides | |
| CN108904519A (en) | Hericium erinaceus Polysaccharides are in the purposes prepared in the drug for treating colitis and include its pharmaceutical composition | |
| CN115300624A (en) | Application of ginsenoside and PD-1 blocker in preparation of head and neck squamous cell carcinoma resisting medicine | |
| CN107519327A (en) | A kind of Phellinus Chinese medicine composition and its extracting method and the application in antineoplastic is prepared | |
| CN102232957B (en) | Use of 3-acetoxyl-8, 24-lanostadiene-21-acid in preparing medicines for preventing or treating liver cancer or breast cancer | |
| TWI551291B (en) | A pharmaceutical or food composition of antrodia cinnamomea extract reducing drug-resistance in cancer cells | |
| CN104906145A (en) | Medical application of paecilomyces hepiali mutant strain PH40 in treating diabetes | |
| CN105012366B (en) | A kind of bright moon grass polysaccharide and its application in preparing for immunological regulation and anti-tumor drug and functional food | |
| Halpern | Medicinal mushrooms | |
| CN103566058B (en) | A kind of double blue or green composition of medicine, preparation method and applications | |
| CN108324707A (en) | Fenofibrate list medicine and combined chemotherapy medicine application in preparation of anti-tumor drugs | |
| CN102793663A (en) | Sustained-release microsphere injection containing antitumor drug (2-methoxyestradiol) | |
| CN101269110B (en) | Radix astragali fermentation liquor for inhibiting cancer cell growth and fermentation method | |
| CN103830262A (en) | Auxiliary drug used for treating cancer, and applications 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 | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |