CN104224857B - A kind of Hericium erinaceus hypha extract is preparing the application in resisting straight colon cancer drug - Google Patents
A kind of Hericium erinaceus hypha extract is preparing the application in resisting straight colon cancer drug Download PDFInfo
- Publication number
- CN104224857B CN104224857B CN201310234812.8A CN201310234812A CN104224857B CN 104224857 B CN104224857 B CN 104224857B CN 201310234812 A CN201310234812 A CN 201310234812A CN 104224857 B CN104224857 B CN 104224857B
- Authority
- CN
- China
- Prior art keywords
- hericium erinaceus
- htj
- weight
- extract
- colon cancer
- 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 hypha extract is preparing the application in resisting straight colon cancer drug.The extract be by Hericium erinaceus mycelium Jing after normal temperature homogenate extraction twice reduced pressure concentration again, last vacuum freeze drying and prepare.The extract has the effect for resisting straight colon cancer, can be used to prepare anti-straight colon cancer drug.
Description
Technical field
The invention belongs to biological pharmacy technical field, and in particular to Hericium erinaceus hypha extract and its preparing anti-straight knot
Application in bowelcancer medicine.
Background technology
Gastrointestinal cancer such as liver cancer, cancer of the stomach, straight colon cancer are one of modal cancers, estimate to account for according to ACS
25%. straight colon cancers of all cancers are one of most common and most cancer types of the death rate in western countries, and straight colon cancer is past
Just it is found toward after 50 years old, M ＆ M occupies cancer the 3rd.Straight colon cancer is less than west in the incidence of disease of China
Fang Guojia, but substantially increase in recent years, particularly in developed regions.The treatment of gastrointestinal cancer include operation, chemotherapy,
Radiotherapy and immunotherapy.Although chemotherapy is most common method thoroughly cannot cure.In in the past few decades, point
The development of sub- biology is made significant progress on straight colon is treated, and the chemotherapy of straight colon cancer is particularly shifted to late period,
Five-year survival rate can be significantly improved, but drug tolerance and toxicity still limit the progress for the treatment of method, it is difficult to be cured.Cause
This, is badly in need of finding and developing the new medicine for resisting straight colon cancer of high-efficiency low-toxicity.
Medicinal fungi is referred to can treat disease, the class fungi with medical value, i.e., in mycelium, fructification, sclerotium
Or such as amino acid, protein, vitamin, polysaccharide and glycoprotein, glucoside, alkaloid, sterols, anthraquinone in spore, can be produced
The multiclass material such as class, flavonoids, has health-care effect to human body, the fungi for having prevention suppression or therapeutic action to disease.Fungi makees
For the medicinal history existing more than 2000 years in China, early in《Sheng Nong's herbal classic》In just describe the fungi-medicines such as Poria cocos, umbellate pore furgus,
《Compendium of Materia Medica》In also recorded more than 20 kinds of biological medicinal fungus, as medicinal and test, current China's tradition confirms that tool is medicative
Fungi is concentrated mainly on load bacterium subphylum and Ascomycotina more than 400 kinds.But it is daily be widely used in clinic but only have
More than 20 plant,《Chinese Pharmacopoeia》Version has clearly included 7 kinds within 2010, and this research of explanation to biological medicinal fungus is still far from perfect, and opens
Send out space larger.Existing research focuses primarily upon a few kind such as glossy ganoderma, Chinese caterpillar fungus, and makees mainly for fungi polysaccharide
For the research and development and application of formulation.
As the development of bioscience, fungi, Acarasiales, lichens etc. are separated from plant kingdom, fungus is individually set up
Boundary.In addition to the glossy ganoderma, Poria cocos, Chinese caterpillar fungus, trametes robinioplila species that have long enjoyed a good reputation at present, the kind of the selected Chinese medicine in the boundary is also less, but wherein
Thousands of kinds of higher fungus, will undoubtedly occur more such fungus medicines from now on, have a high potential, it will greatly enrich Chinese medicine treasure-house.
Additionally, the fungus medicines based on medicinal fungi, in addition to tradition is using fructification, the modern times have developed to adopt liquid（Deep layer）Send out
Ferment（Application bacterium ball and zymotic fluid）, solid fermentation（Application mycoplasma）Etc. production technology.Solid fermentation produce common mycoplasma its master
Want medicinal part to be still the mycelium and its secondary substance contained by it, but it should be noted that medicinal fungus fructification removes itself
It is used as medicine outer, is also used for Chinese traditional medicine biology engineering research, to develop new drug.And Fermentation Engineering, especially China are distinctive solid
Body Fermentation Engineering, the time that it is present existing more than 2000 years history, by prolonged solid fermentation, generate biological metabolism product
Thing includes biological enzyme material, and the medicinal mycoplasma which generates is than liquid fermentation good drug efficacy.But solid fermentation existing defects, it is easily miscellaneous
Bacterium pollutes, and existing solid-fermented technique research is studies in China focus, and the direction of main research is sterile culture, second incubation, miscellaneous
Bacterium suppresses, Chinese medicine implements two-way solid-fermented technique as Nutrient medium, breaks through the bottleneck of solid-fermented technique, realizes biological medicament
With the extensive industrialization of bacterium.
Hericium erinaceus is a kind of hedgehog hydnum Cordycepps mushroom（Fungi）, had 2000 year for treating enterogastric diseases in Chinese medicine
History, a series of micromolecular compounds such as erinacines for containing are considered to have nerve regneration, and can pass through blood
Brain barrier repairs damaged nerve tissue, and Hericium erinaceus also has antiulcer, anti-inflammatory, and antimicrobial, immunological regulation, improves liver function,
Anti-aging, reduces blood sugar and blood fat, improves anti-anoxia ability, increases heart output, improves the effects such as remarks circulation, comprising hedgehog hydnum
The medical preparation of bacterium is widely used in China.
Above all polysaccharide and glycoprotein in the mycelial active material of Hericium erinaceus, at present both at home and abroad to Hericium erinaceus polysaccharide
Research shows that Hericium erinaceus Polysaccharides have multiple biological activities and pharmacological action, can strengthen the phagocytic function of macrophage, promotes molten
The formation of sanguinin, anti-leucocyte decline, and hypoglycemic, anticoagulation, antithrombotic, anti-mutation and anti-ageing wait for a long time.Therefore, Hericium erinaceus Polysaccharides
Enjoy people to pay close attention to, become the focus of the area research such as molecular biology, medicine, Food Science and development and application in recent years.《Lake
South saves Chinese medicine standard》Version in 2009,122-123 page describes Hericium erinaceus culture, Hericium erinaceus mycelium of the present invention
It is the Hericium erinaceus culture of standard record.
Currently, screened from higher fungus and its culture and found active component, be applied to antitumor, disease-resistant
Poison, and treatment diabetes etc., not only become the developing direction that state key studies new drug, and countries in the world today exploitation is biological
The focus of medicine.But, at present and have no and be applied to resist the report of straight colon cancer by Hericium erinaceus mycelial extract.
The content of the invention
It is contemplated that overcoming the deficiencies in the prior art, there is provided a kind of Hericium erinaceus hypha extract, the Hericium erinaceus mycelia
Body extract can kill people's carcinoma of the rectum SW620 and HT-29 cells in vitro and can suppress the growth of carcinoma of the rectum transplantable tumor in vivo,
Therefore can be applicable to prepare and resist straight colon cancer drug.
The preparation method of Hericium erinaceus hypha extract of the present invention comprises the steps：
（1）Hericium erinaceus mycelium is taken, first time water normal temperature homogenate extraction is carried out, is extracted 15～25 minutes, then carry out the
Secondary water normal temperature homogenate extraction, extracts 10～20 minutes, and in first time water normal temperature homogenate extraction, the weight of water is Hericium erinaceus mycelia
4～6 times of body weight, preferably 5 times；In second water normal temperature homogenate extraction, the weight of water is the 2 of Hericium erinaceus mycelium weight
～4 times, preferably 3 times；
（2）The supernatant obtained after extract is centrifuged has been evaporated to Precipitation under the conditions of 55～60 DEG C；Will
Concentrate vacuum freeze drying under the conditions of -60～-55 DEG C, had both obtained Hericium erinaceus hypha extract.
Wherein, the temperature of the water normal temperature homogenate extraction is 20～50 DEG C.Step（2）Described in extract centrifugal condition
For：3500～4500 revs/min of rotating speed, is centrifuged 8～15 minutes.
Preferably, the preparation process of the extract also includes：By step（2）Concentrate again with weight be the concentrate
4～6 times of weight and concentration of volume percent be 70～80% ethanol dissolving, will dissolving part in 55～60 DEG C of reduced pressure concentrations
To without alcohol taste, by gained concentrate in -60～-55 DEG C of vacuum freeze dryings.
Preferably, the preparation process of the extract also includes：By step（2）Concentrate again with weight be the concentrate
4～6 times of weight and concentration of volume percent be 70～80% ethanol dissolving, by insoluble part in -60～-55 DEG C of vacuum
Freeze-drying.
Hericium erinaceus mycelium of the present invention is《Hunan Province's Chinese medicine standard》Version in 2009, the monkey of 122-123 page of record
Head bacterium culture, the Hericium erinaceus mycelium are hedgehog hydnum Cordycepps fungi Hericium erinaceus Hericium erinaceum（Bull.ex Fr.）
The drying composite of the solid medium that mycelium Pers. is grown nonparasitically upon another plant with which.
Normal temperature homogenate extraction Hericium erinaceus mycelium of the present invention, under normal temperature or room temperature（20-50℃）, do not heat, add water
Power beating formula it is water-soluble go out active ingredient, without temperature damage, can dissolve active ingredient and not destroy active ingredient；60 DEG C of decompressions
Concentration, will not destroy active ingredient, particularly active enzyme material；In order to not affect relevant active material, present invention employs
Vacuum freeze drying means.
The present invention has carried out straight colon cancer SW620 of suppression people and HT-29 cells to the Hericium erinaceus hypha extract of gained
And animal resists the experiment of straight colon cancer transplantable tumor, knowable to the result of Fig. 1 to Fig. 9 and Biao 1 to table 3, the Hericium erinaceus mycelium is carried
Take thing and there is the effect for resisting straight colon cancer, can be used to prepare anti-straight colon cancer drug.
Description of the drawings
Fig. 1:Inhibitory action of the HTJ#2 to the straight colon cancer SW620 cell of people.Concentration be 0.078-2.5 mg/ml, 72
Hour drug-treated, Thiazolyl blue（MTT）Method determines cytoactive；
Fig. 2:Inhibitory action of the HTJ#5 to the straight colon cancer SW620 cell of people.Concentration be 0.39-12.5 mg/ml, 72
Hour drug-treated, Thiazolyl blue（MTT）Method determines cytoactive；
Fig. 3:Inhibitory action of the HTJ#5 to the straight colon cancer cell line HT-29 of people.Concentration be 0.3125-20 mg/ml, 72
Hour drug-treated, Thiazolyl blue（MTT）Method determines cytoactive；
Fig. 4:Inhibitory action of the HTJ#5A to the straight colon cancer cell line HT-29 of people.Concentration be 0.3125-20 mg/ml, 72
Hour drug-treated, Thiazolyl blue（MTT）Method determines cytoactive；
Fig. 5:Fig. 5 A are HTJ#2, the curative effect point in HTJ#5 and 5-Fluorouracil body to the straight colon cancer SW620 transplantable tumor of people
Analysis；Fig. 5 B are HTJ#2, the animal toxicity of the straight colon cancer SW620 transplantable tumor of people are analyzed in HTJ#5 and 5-Fluorouracil body；It is right
According to group, 5-Fluorouracil（35 mg kg days）, HTJ#2 (200 or 500 mg kg day), and HTJ#5 (200 or 500
Mg kg day), it is by continuous 5 days lumbar injections or oral administration, little to the SCID of the straight colon cancer SW620 transplantable tumor of lotus people
The Synergistic action and toxicity of mouse.Every group of 3-5 animal；
Fig. 6:Fig. 6 A are HTJ#5, the curative effect point in HTJ#5A and 5-Fluorouracil body to the straight colon cancer HT-29 transplantable tumor of people
Analysis；Fig. 6 B are HTJ#5, the animal toxicity of the straight colon cancer HT-29 transplantable tumor of people are analyzed in HTJ#5A and 5-Fluorouracil body.It is right
According to group, 5-Fluorouracil（30 mg kg days, lumbar injection）, HTJ#5 (500 mg kg days, oral), HTJ#5
(500 mg kg days, oral), are administered, per group of 5 animals for continuous 5 days；
Fig. 7:HTJ#5, HTJ#5A, and individual curative effect of the 5-Fluorouracil abdominal cavity to the straight colon cancer HT-29 transplantable tumor of people；It is right
According to group, 5-Fluorouracil（30 mg kg days, lumbar injection）, HTJ#5 (500 mg kg days, oral), and HTJ#5A
(500 mg kg days, oral), were administered by continuous 5 days；Wherein, Fig. 7 A are control group, and Fig. 7 B are 5-Fluorouracil（30
Mg kg day, lumbar injection）, Fig. 7 C are HTJ#5 (500 mg kg days, oral), and Fig. 7 D are HTJ#5A (500 millis
G/kg/day, oral)；In figure, abscissa is the time（My god）, ordinate is tumor weight（Milligram）；
Fig. 8:Fig. 8 A are HTJ#5, the curative effect point in HTJ#5A and 5-Fluorouracil body to the straight colon cancer HT-29 transplantable tumor of people
Analysis；Fig. 8 B are HTJ#5, the animal toxicity of the straight colon cancer HT-29 transplantable tumor of people are analyzed in HTJ#5A and 5-Fluorouracil body；It is right
According to group, 5-Fluorouracil（30 mg kg days, lumbar injection）, HTJ#5 (1000 mg kg days, oral), HTJ#5
(1000 mg kg days, oral), are administered for continuous 5 days. per group of 5 animals；
Fig. 9:HTJ#5, HTJ#5A, and individual curative effect of the 5-Fluorouracil abdominal cavity to the straight colon cancer HT-29 transplantable tumor of people;It is right
According to group, 5-Fluorouracil（30 mg kg days, lumbar injection）, HTJ#5 (1000 mg kg days, oral), and HTJ#
5A (1000 mg kg days, oral), was administered by continuous 5 days;Wherein, Fig. 9 A are control group, and Fig. 9 B are 5-Fluorouracil
（30 mg kg days, lumbar injection）, Fig. 9 C are HTJ#5 (1000 mg kg days, oral), and Fig. 9 D are HTJ#5A
(1000 mg kg days, oral)；In figure, abscissa is the time（My god）, ordinate is tumor weight（Milligram）.
Specific embodiment
The present invention is further illustrated with reference to the accompanying drawings and examples.
Embodiment 1
A kind of Hericium erinaceus hypha extract, its preparation method comprise the steps：
（1）Hericium erinaceus mycelium is taken, first time water normal temperature homogenate extraction is carried out, is extracted 20 minutes, then carry out second
Water normal temperature homogenate extraction, extracts 15 minutes；In first time water normal temperature homogenate extraction, the weight of water is Hericium erinaceus mycelium weight
5 times；In second water normal temperature homogenate extraction, the weight of water is 3 times of Hericium erinaceus mycelium weight；
（2）The supernatant obtained after the extract centrifugation that will be obtained after above-mentioned extraction twice reduced pressure concentration under the conditions of 60 DEG C
Concentrate is obtained to there is Precipitation；By concentrate under the conditions of -60 DEG C vacuum freeze drying, obtain final product Hericium erinaceus mycelium extraction
Thing.
Wherein, the temperature of the water normal temperature homogenate extraction is 30 DEG C.Step（2）Described in extract centrifugal condition be：Turn
4000 revs/min of speed, is centrifuged 10 minutes.
Embodiment 2
A kind of Hericium erinaceus hypha extract, its preparation method comprise the steps：
（1）Hericium erinaceus mycelium is taken, first time water normal temperature homogenate extraction is carried out, is extracted 20 minutes, then carry out second
Water normal temperature homogenate extraction, extracts 15 minutes；In first time water normal temperature homogenate extraction, the weight of water is Hericium erinaceus mycelium weight
5 times；In second water normal temperature homogenate extraction, the weight of water is 3 times of Hericium erinaceus mycelium weight；
（2）The supernatant obtained after the extract centrifugation that will be obtained after above-mentioned extraction twice reduced pressure concentration under the conditions of 60 DEG C
Concentrate is obtained to there is Precipitation；
（3）By step（2）Concentrate again with weight be 5 times of the concentrate weight and concentration of volume percent be 80%
Ethanol dissolving, dissolving part is evaporated to without alcohol taste at 60 DEG C, by gained concentrate in -60 DEG C of vacuum freeze dryings,
Obtain the Hericium erinaceus hypha extract for further extracting.
Wherein, the temperature of the water normal temperature homogenate extraction is 30 DEG C.Step（2）Described in extract centrifugal condition be：Turn
4000 revs/min of speed, is centrifuged 10 minutes.
Embodiment 3
A kind of Hericium erinaceus hypha extract, its preparation method comprise the steps：
（1）Hericium erinaceus mycelium is taken, first time water normal temperature homogenate extraction is carried out, is extracted 20 minutes, then carry out second
Water normal temperature homogenate extraction, extracts 15 minutes；In first time water normal temperature homogenate extraction, the weight of water is Hericium erinaceus mycelium weight
5 times；In second water normal temperature homogenate extraction, the weight of water is 3 times of Hericium erinaceus mycelium weight；
（2）The supernatant obtained after the extract centrifugation that will be obtained after above-mentioned extraction twice reduced pressure concentration under the conditions of 60 DEG C
Concentrate is obtained to there is Precipitation；
（3）By step（2）Concentrate again with weight be 5 times of the concentrate weight and concentration of volume percent be 80%
Ethanol dissolving, by insoluble part in -60 DEG C of vacuum freeze dryings, obtain the Hericium erinaceus hypha extract for further extracting.
Wherein, the temperature of the water normal temperature homogenate extraction is 30 DEG C.Step（2）Described in extract centrifugal condition be：Turn
4000 revs/min of speed, is centrifuged 10 minutes.
Embodiment 4
Hericium erinaceus hypha extract Functional Validation Test-to colorectal cancer SW620 and the inhibitory action of HT-29 cells
First, materials and methods
1st, cell and culture:Straight colon cancer SW620 of people and HT-29 cells are from American Type Culture research institute
(American Type Culture Collection,ATCC;Manassas,VA,UAS).SW620 cells raise in
RPMI1640 nutrient solutions, HT-29 cells are raised in DMEM nutrient solutions, containing 10% calf serum（Atlanta Biological,
Lawrenceville, GA, USA）, the penicillin of 100 units per mls, and 0.1 mcg/ml streptomysin (Invitrogen,
Grand Island, NY, USA).
2nd, medicine：The Hericium erinaceus hypha extract finally obtained by 1 preparation method of embodiment, is named as HTJ#2；By reality
The Hericium erinaceus hypha extract that 2 preparation method of example is finally obtained is applied, HTJ#5 is named as；Finally obtained by 3 preparation method of embodiment
The Hericium erinaceus hypha extract for obtaining, is named as HTJ#5A；Aforementioned Hericium erinaceus hypha extract is dissolved in respectively containing 0.1%
In RPMI1640 (SW620 cells) DMEM (HT-29 cells) nutrient solution of DMSO.
3rd, drug concentration and scheme:HTJ#2, HTJ#5, and HTJ#5A drug concentration is in 0.625-20mg/ml（Milligram/milli
Rise）Continuous 72 hours drug-treateds.
4. Human colorectal carcinoma SW620 and HT-29 cells growth activity is suppressed：72 hours drug-treateds, then use Thiazolyl blue
(MTT) method determines cytoactive.
2nd, experimental result
Growth inhibition effects of the 1.HTJ#2 and HTJ#5 to the straight colon cancer SW620 cell of people.
The straight colon cancer SW620 cell 72 of people that we process in vitro culture with eight kinds of different HTJs (#1-#8) first is little
When observing the growth in vitro inhibitory action to straight colon cancer cell.Result of the test shows that all HTJs can effectively suppress people
The growth of straight colon cancer SW620 cell, its inhibitory action amount effect curve（Data do not show）.Wherein, HTJ#2 and HTJ#5
Inhibitory action is stronger.Suppression of Fig. 1 as shown by data HTJ#2 to the straight colon cancer SW620 cell of people shows and makes of very strong, its half
Casualty-producing concentrations (IC50) for 0.8 mg/ml, HTJ#2 can kill the SW620 cells more than 85% in 1.25 mg/mls.Fig. 2
Half casualty-producing concentrations (IC of as shown by data HTJ#5 to the straight colon cancer SW620 cell of people50) for 2.0 mg/mls, HTJ#5 exists
6.25 mg/mls can kill the SW620 cells more than 85%.
Growth inhibition effects of the 2.HTJ#5 and HTJ#5A to the straight colon cancer cell line HT-29 of people.
In general, the straight colon cancer cell line HT-29 of people compares resistance to most chemotherapy.We use HTJ#5 and HTJ#5A72
Hour the HT-29 cells of culture are processed observing its growth in vitro inhibitory action.Test data is shown in Fig. 3 to Fig. 4.Data result table
The bright process by 72 hours HTJ#5 or HTJ#5A to cell can effectively suppress the growth of HT-29 cells, its inhibitory action amount
Effect curve.Half casualty-producing concentrations (IC of Fig. 3 as shown by data HTJ#5 to HT-29 cells50) for 1.25 mg/mls, HTJ#5 exists
20 mg/mls can kill the HT-29 cells more than 80%.Fig. 4 as shown by data HTJ#5A to HT-29 cellkilling capacities with
HTJ#5 is similar or stronger.Its half casualty-producing concentrations (IC50) for 1.25 mg/mls, HTJ#5A can be killed in 20 mg/mls
HT-29 cells more than 90%.
Embodiment 5
Hericium erinaceus hypha extract Functional Validation Test
First, materials and methods
1st, animal：The SCID raettins of 8～12 weeks（18～22g of weight）, from Roswell Park Cancer
Institute (Buffalo, NY), and raise by regulation.
2nd, medicine：The Hericium erinaceus hypha extract finally obtained by 1 preparation method of embodiment, is named as HTJ#2；By reality
The Hericium erinaceus hypha extract that 2 preparation method of example is finally obtained is applied, HTJ#5 is named as；Finally obtained by 3 preparation method of embodiment
The Hericium erinaceus hypha extract for obtaining, is named as HTJ#5A；Aforementioned Hericium erinaceus hypha extract is dissolved in into the nothing containing 10%DMSO
In bacterium physiological saline, 5-Fluorouracil（5-FU：50mg/ml）Hoffmann-La Roche are purchased from, Inc (Nutley, NJ) is with nothing
Bacterium normal saline dilution..
3rd, tumour cell：The straight colon cancer cells of SW620 and HT-29.About 1000000 trainings are injected in the foundation of transplantable tumor first
Foster cell is simultaneously transplanted through number generation.
4th, drug dose and scheme：HTJ#2 is oral or injects 200-500 mg kg days, and HTJ#5 is oral or injects
200-1000 mg kg days, the oral 500-1000 mg kg days of HTJ#5A, 30-35 mg/kg of 5-Fluorouracil/
My god, intraperitoneal injection, once a day, the course for the treatment of 5 days.Control group is oral or injects 10%DMSO solution, and mouse is given per 20g body weight
Give 200-400 μ l（It is identical with experimental group）.Every group of 3-5 mouse, in part Experiment, the two kinds of different transplanting of same mouse lotus
Knurl.
5. measurement of tumor：With the axis of vernier caliper measurement tumour（L represents major axis, and W represents short axle）Tumor weight is pressed
Formula is calculated：Tumor weight=1/2 (L x W2) is (mm). and (1mg=1mm3), the daily same time carries out measurement of tumor, weekly 3-4
It is secondary.
6. maximum tolerated dose（MTD）And toxicity assessment：Maximum tolerated dose is defined as administration causes Mouse Weight to mitigate
20% or so, it is not result in that medicine causes the dosage of dead mouse.Receive front ten days after drug therapy, measure tumour daily
And Mouse Weight, followed by measured once per two days.
7. antitumor activity：Antitumor activity evaluation index, maximum Tumor growth inhibition, with the average tumor weight for the treatment of group
The exemplary embodiment lock of amount and control group is calculating；Tumor doubling time, is that tumor weight rises to the two of initial weight
Average time needed for times；Tumor section reacts (PR), when tumor size reduces half（To initial 50% or more）When；It is swollen
Knurl reacts (CR) completely, when cannot detect tumour（Cases of complete remission）When；Healing is defined as at least 60 days after drug therapy
Tumour is can't detect inside（Reach tumour and react holding at least 60 days completely, now general tumour will not recur, and animal no longer protects
Stay）.
2nd, experimental result
1st, HTJ#2, HTJ#5 and 5-Fluorouracil are antitumor to the SCID mice of the straight colon cancer transplantable tumor of lotus SW620 people
Activity and toxicity.
Our external MTT tests find that HTJ#2 and HTJ#5 has obvious cell to the straight colon cancer SW620 cell of people
Toxicity.The IC of HTJ#2 and HTJ#550It is 0.8mg/ml and 2.0mg/ml respectively.Therefore HTJ#2 and HTJ#5 be we used in lotus
The SCID mice of SW620 transplantable tumors is observing its antitumor activity.The test data of Fig. 5 and Biao 1 shows vehicle control group, five fluorine
Uracil dosage is 35 mg kg days, HTJ#2 and HTJ#5 dosage be 200 mg kg days and 500 mgs/kg/
My god, once a day, the antitumor activity and toxicity of the SCID mice to the straight colon cancers of lotus SW620 of continuous 5 days.Result of the test table
Bright SW620 growth of transplanted human is very fast, and mean doubling time is 3.1 ± 0.1.HTJ#2 and HTJ#5 is used in oral and lumbar injection
When dosage is 200 mg kg days and 500 mg kg day, for tumor suppression and prolongation TDT have one
Fixed effect.However, HTJ#2 shows overt toxicity in 500 mg kg day lumbar injections, and 67% test is caused to be moved
Thing death (2/3rds animal deads).And same dose of HTJ#2（500 mg kg days, when oral, without substantially poison
Property.. data also show 5-Fluorouracil in 35 mg kg days, and lumbar injection only has 48.4% suppression for SW620 transplantable tumors
Rate processed, and extend TDT from 3.1 ± 0.1 days（Vehicle control group）By 4.8 ± 0.4 days.However, 5-Fluorouracil
In 35 mg kg days) toxicity is too big and cause 22.5% to lose weight, maximum tolerated dose can be can exceed that.Data display
HTJ#2 is similar compared to 5-Fluorouracil anti-tumor activity with HTJ#5 or higher（Inhibiting rate 48.7-61.4%）Toxicity is low, but extends
TDT is shorter（～five days）Show that tumour recovers very fast after drug therapy stopping.As a result also indicate that HJT#2 and
HTJ#5 is orally similar to the anti-knurl curative effect of lumbar injection.
2.HTJ#5, HTJ#5A and 5-Fluorouracil antitumor activity and poison to the SCID mice of lotus HT-29 transplantable tumors
Property.
According to the experience before us, SW620 transplantable tumors are more more sensitive than HT-29 transplantable tumor for most of chemicalses.
So, we apply HTJ#5, HTJ#5A and 5-Fluorouracil to the SCID mice of the straight colon cancer tumours transplantable tumors of lotus HT-29
Antitumor activity and toxicity.Result of the test is shown in that figure .6-7 and the test data of table 2 show vehicle control group, 5-Fluorouracil dosage
For 30 mg kg days, lumbar injection（Due to 35 mg kg days, toxicity is too big）, HTJ#5 and HTJ#5A dosage is 500
Mg kg day, orally, once a day, continuous 5 days.As shown by data, HT-29 doubling times are 8.7 ± 1.0 days, five fluorine urine
The tumor control rate of pyrimidine is 42.7 ± 10%.However, and being obviously prolonged TDT to 14.2 days (P<0.05), orally
500 mg kg day of HTJ#5 dosage, treats 5 days, and to HT-29 transplantable tumors, inhibition rate of tumor growth is 65.0 ± 17.3,
Doubling time reaches 19.3 ± 4.9 days, more higher than 5-Fluorouracil dosage anti-tumor activity, and toxicity is substantially low.Oral HTJ#5A500
Mg kg day, the course for the treatment of 5 days, Tumor growth inhibition extend to 19.2 ± 3.3 days for 66.2 ± 17.8 doubling times, also than five
Fluorouracil dosage anti-tumor activity is higher, and toxicity is substantially low.
We improve HTJ#5 and HTJ#5A again to 1000 mg kg days, see whether that can enter one improves anti-knurl treatment
Effect.Result of the test is shown in figure .8-9 and table 3.As shown by data, HT-29 doubling times are 6.4 ± 0.3 days, than last time growth of transplanted human
Comparatively fast.The tumor control rate of 5-Fluorouracil is improved to 63.2 ± 5.9%.However, extending TDT to 10.4 days (P<
0.05), oral 1000 mg kg day of HTJ#5 dosage, treats 5 days, and to HT-29 transplantable tumors, inhibition rate of tumor growth is
66.8 ± 4.6, the doubling time reaches 16.2 ± 1.3 days, more higher than 5-Fluorouracil dosage anti-tumor activity, and toxicity is substantially low.Mouthful
Take HTJ#5A1000 mg kg days, the course for the treatment of 5 days, Tumor growth inhibition extends to 15.6 for 64.2 ± 2.9 doubling times ±
1.7 days, also higher than 5-Fluorouracil dosage anti-tumor activity, toxicity was substantially low.As a result show improve HTJ#5 and HTJ#5A from
500 mg kg days do not have into one and improve anti-knurl curative effect, do not increase toxicity yet to 1000 mg kg days.
Conclusion：
1.HTJ#2 and HTJ#5 have obvious inhibitory action to the straight colon cancer SW620 cell of people in vitro.
2.HTJ#5 and HTJ#5A have obvious inhibitory action to the straight colon cancer cell line HT-29 of people in vitro.
3.HTJ#2 and HTJ#5 is in 200 or 500 mg kg days）In vivo to the straight colon cancer SW620 department of human head and neck of people
Cancer transplantable tumor has certain antitumor curative effect, similar or higher to 5-Fluorouracil dosage anti-tumor activity, but extends tumour multiplication
Time is shorter.
4.HJT#2 and HTJ#5 are orally similar to the anti-knurl curative effect of lumbar injection.
5.HTJ#2 has overt toxicity but oral no overt toxicity in 500 mg kg day lumbar injections.
6.HTJ#5 and HTJ#5A（500-1000 mg kg days, oral 5 days）Compare 5-Fluorouracil（30-35 milligrams/
Kg/day, lumbar injection 5 days）Anti- knurl good effect to HT-29 transplantable tumors and toxicity is substantially low.
7. result shows to improve HTJ#5 and HTJ#5A from 500 mg kg days to 1000 mg kg days, it is impossible to enter
One is improved anti-knurl curative effect, does not also increase toxicity.
Table 1:Summarize HTJ#2, HTJ#5 and 5-Fluorouracil anti-knurl curative effect and animal toxicity to SW620 colorectal cancers
Control group is given 0.2 milliliter of physiological saline daily containing 10% DMSO by mouse body weight per 20 grams.Each experimental group
Using 5 mouse.Transplantable tumor transplanting starts treatment after reaching about 190 to 220 milligrams within 7 days.
Table 2:Summarize HTJ#5, HTJ#5A and anti-knurl curative effect and animal toxicity with 5-Fluorouracil to the straight colon cancers of HT-29
Control group is given 0.2 milliliter of physiological saline daily containing 10% DMSO by mouse body weight per 20 grams.Each experimental group
Using 5 mouse.Transplantable tumor transplanting starts treatment after reaching about 180 to 200 milligrams within 7 days.
Table 3:Summarize HTJ#5, HTJ#5A and anti-knurl curative effect and animal toxicity with 5-Fluorouracil to the straight colon cancers of HT-29
Control group is given 0.2 milliliter of physiological saline daily containing 10% DMSO by mouse body weight per 20 grams.Each experimental group
Using 5 mouse.Transplantable tumor transplanting starts treatment after reaching about 200 to 230 milligrams within 7 days.
Claims (2)
1. a kind of Hericium erinaceus hypha extract is preparing the application in resisting straight colon cancer drug；Characterized in that, the hedgehog hydnum
The preparation method of bacterium hypha extract comprises the steps：
（1）Hericium erinaceus mycelium is taken, first time water normal temperature homogenate extraction is carried out, is extracted 15～25 minutes, then carry out second
Water normal temperature homogenate extraction, extracts 10～20 minutes；In first time water normal temperature homogenate extraction, the weight of water is Hericium erinaceus mycelia body weight
4～6 times of amount；In second water normal temperature homogenate extraction, the weight of water is 2～4 times of Hericium erinaceus mycelium weight；The hedgehog hydnum
Bacterium mycelium is hedgehog hydnum Cordycepps fungi Hericium erinaceus Hericium erinaceum（Bull.ex Fr.）Pers. mycelium and its
The drying composite of the solid medium grown nonparasitically upon another plant；The temperature of the water normal temperature homogenate extraction is 20～50 DEG C；
（2）The supernatant obtained after the extract centrifugation that will be obtained after above-mentioned extraction twice reduced pressure concentration under the conditions of 55～60 DEG C
Concentrate is obtained to there is Precipitation；By concentrate under the conditions of -60～-55 DEG C vacuum freeze drying, obtain final product Hericium erinaceus mycelia
Body extract；The extract centrifugal condition is：3500～4500 revs/min of rotating speed, is centrifuged 8～15 minutes；
（3）By step（2）Concentrate again with weight be 4～6 times of the concentrate weight and concentration of volume percent be 70～
80% ethanol dissolving, dissolving part is evaporated to without alcohol taste at 55～60 DEG C, and gained concentrate is true at -60～-55 DEG C
Vacuum freecing-dry.
2. it is as claimed in claim 1 to apply, it is characterised in that in Hericium erinaceus hypha extract preparation method, first time water is normal
In warm homogenate extraction, the weight of water is 5 times of Hericium erinaceus mycelium weight；In second water normal temperature homogenate extraction, the weight of water
For 3 times of Hericium erinaceus mycelium weight.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310234812.8A CN104224857B (en) | 2013-06-14 | 2013-06-14 | A kind of Hericium erinaceus hypha extract is preparing the application in resisting straight colon cancer drug |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310234812.8A CN104224857B (en) | 2013-06-14 | 2013-06-14 | A kind of Hericium erinaceus hypha extract is preparing the application in resisting straight colon cancer drug |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104224857A CN104224857A (en) | 2014-12-24 |
| CN104224857B true CN104224857B (en) | 2017-03-29 |
Family
ID=52214336
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310234812.8A Active CN104224857B (en) | 2013-06-14 | 2013-06-14 | A kind of Hericium erinaceus hypha extract is preparing the application in resisting straight colon cancer drug |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104224857B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113730448B (en) * | 2021-09-06 | 2022-10-18 | 天津市农业科学院 | Preparation method of hericium erinaceus anti-human colon cancer tumor cell (HCT-8) extract |
Family Cites Families (3)
| 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 |
| CN104224856B (en) * | 2013-06-06 | 2017-04-19 | 湖南新汇制药股份有限公司 | Hericium erinaceus mycelium extract and application thereof in preparation of head-neck cancer resistant drugs |
| CN104224855A (en) * | 2013-06-06 | 2014-12-24 | 湖南新汇制药股份有限公司 | Use of hericium erinaceus hyphae extract in preparation of drug for resisting stomach cancer |
-
2013
- 2013-06-14 CN CN201310234812.8A patent/CN104224857B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN104224857A (en) | 2014-12-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102153668B (en) | Anticancer Armillaria luteovirens polysaccharide and extraction process thereof | |
| CN103446385A (en) | Traditional Chinese medicine preparation for treating diabetic complications | |
| CN104224855A (en) | Use of hericium erinaceus hyphae extract in preparation of drug for resisting stomach cancer | |
| CN104224857B (en) | A kind of Hericium erinaceus hypha extract is preparing the application in resisting straight colon cancer drug | |
| Stengler | Health benefits of medicinal mushrooms | |
| CN108570116B (en) | Pleurotus citrinopileatus polysaccharide, preparation method and medical application in preventing and treating diabetes | |
| CN103720722A (en) | Application of water-soluble lachnum melanin in preparing medicine for inhibiting liver cancer | |
| CN104224856B (en) | Hericium erinaceus mycelium extract and application thereof in preparation of head-neck cancer resistant drugs | |
| CN102441000B (en) | Anti-cancer anti-disease buccal tablet | |
| CN104248647B (en) | A kind of Hericium erinaceus (Bull. Ex Fr.) Pers. mycelium extract and preparing the application in medicines resistant to liver cancer | |
| CN102078600B (en) | Anti-cancer compound ganoderma composition, application thereof and pharmaceutical composition containing same | |
| CN103230418B (en) | Mycelium extract for helicobacter pylori-resistant biological medicine and preparation technology thereof | |
| US11065293B2 (en) | Pharmaceutical composition for decreasing the side effects of cancer drug, and manufacturing method and uses thereof | |
| CN105535011A (en) | Applications of mushroom mycelium polysaccharides | |
| CN107893035A (en) | A kind of culture medium of Hericium erinaceus, sealwort bioconversion mycelium, the mycelial extract of sealwort bioconversion and application thereof | |
| CN107519327A (en) | A kind of Phellinus Chinese medicine composition and its extracting method and the application in antineoplastic is prepared | |
| CN108721532B (en) | For assisting in the treatment of the Chinese medicine composition of lung cancer | |
| CN102106875A (en) | Method for extracting water-soluble alpha-glucosidase inhibitor | |
| CN102423384B (en) | Traditional Chinese drug preparation for treating lung cancer, and preparation method thereof | |
| CN101244041B (en) | Medicament for preventing and treating acute liver damnification and preparation thereof | |
| CN100355415C (en) | Frozen dry powder injection of astrogalus root and its preparing method | |
| CN1284848C (en) | Dry red persimmon wine and its brewing method | |
| CN104857422B (en) | It treats the composition of colorectal cancer and is used to prepare the purposes for the treatment of large intestine cancer drug | |
| CN105288278A (en) | Anti-tumor traditional Chinese medicine microbial fermentation agent and preparation method and application thereof | |
| CN107137567A (en) | Cherry freeze-dried powder freezes compound powder and the application in anti-gout drugs are prepared with smilax |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |