CN108210493B - Pharmaceutical composition for improving gastric precancerous lesion and preparation thereof - Google Patents
Pharmaceutical composition for improving gastric precancerous lesion and preparation thereof Download PDFInfo
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- CN108210493B CN108210493B CN201810201158.3A CN201810201158A CN108210493B CN 108210493 B CN108210493 B CN 108210493B CN 201810201158 A CN201810201158 A CN 201810201158A CN 108210493 B CN108210493 B CN 108210493B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
- A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/12—Ketones
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
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Abstract
The present invention relates to a kind of pharmaceutical composition for improving gastric precancerous lesion and preparation thereof, belong to drug field.Described pharmaceutical composition comprising: Bergenin, Berberine hydrochloride and curcumin.The weight proportion of the Bergenin, Berberine hydrochloride and curcumin is 1-10:1-10:1-10.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition for improving gastric precancerous lesion and preparation thereof, belong to drug field.
Background technique
Gastric cancer is the most common malignant tumour in China, in recent years due to male lung cancer, women with breast cancer incidence sharply on
It rises, causes incidence gastric cancer rate to be down to third from first place, but actual death toll does not subtract, annual death rate has accounted for the world total
40%, more it is disturbing that age of onset was down to 25~35 years old crowd from past 50 years old or more, this group of crowd's disease incidence is also
Increased with speed at double.
Gastric cancer can be divided into gastric pattern and visible peristalsis visible intestinal peristalsis two major classes from tissue generation, be developed to chronic wither by chronic superficial gastritis
Contracting gastritis, mucomembranous surface and adenomatosis and intestinal metaplasia are not true to type or dysplasia, until the process of canceration is typical
Intestinal-type gastric cancer is also the most common type in China.Therefore clinically often companion intestinal metaplasia (IM), dysplasia (Dys) are referred to as
For gastric precancerous lesion (PLGC).It is reported that the canceration rate of external atrophic gastritis is 8.6%-13.8%, country's report is
1.2%-7.1%, the canceration rate of intestinal metaplasia follow-up 10 years are 1.7%, and the canceration rate of slight dysplasia is 2.53%;In
The canceration rate for spending dysplasia is 4%-8%;The canceration rate of severe dysplasia is 10%-83%.Disease has before Stomach Carcinomas
The possibility that effect treatment can reduce gastric cancer occurs, and the discovery rate of early carcinoma is improved, to greatly improve the survival rate of Patients with Gastric Cancer
And existence.
It at this stage, include eliminating pylorus, supplement antioxidant and micro- for the western medical treatment of gastric precancerous lesion
Secondary element, gastric mucosa protectant, antibiotic etc..But for the patient with Helicobacter pylori infection, even if killing pylorus
After the infection of pylori, development of the PLGC to gastric cancer still can not be effectively prevented, and the drugs such as antioxidant and vitamin
It still cannot get preferable data to the therapeutic effect of gastric precancerous lesion to support, it has been determined that effective gastric mucosa protective agent,
It is only capable of slowing down the development process of gastric precancerous lesion, but can not prevent or reverse entire pathologic process.Therefore gastric precancerous lesion
Treatment is still so far the treatment difficult point in doctor trained in Western medicine field.
Currently, western medicine still lacks ideal method to the treatment of gastric precancerous lesion, and traditional Chinese medicine then has one to the treatment of the disease
Determine advantage, achieves good curative effect, it is apparent pre- that a large amount of clinical treatments and experimental study all show that traditional Chinese medicine has PLGC
Anti- even reverse effect.But that there are dosages is big for Chinese medicine tradition prescription, needs patient voluntarily to decoct, quality of medicinal material is unstable to be caused
The shortcomings that uncertain therapeutic efficacy is cut.Although modern Chinese herbal medicine preparation can partially overcome disadvantages associated, the tired of quality of medicinal material is nevertheless suffered from
It disturbs, and effective component is indefinite, is unfavorable for subsequent market exploitation.
Summary of the invention
The first aspect of the present invention is to provide a kind of pharmaceutical composition prevented or treat gastric precancerous lesion comprising: rock
Chinese cabbage element, Berberine hydrochloride and curcumin.
In one embodiment, the weight proportion of the Bergenin, Berberine hydrochloride and curcumin is 1-10:1-
10:1-10。
In another embodiment, the weight proportion of the Bergenin, Berberine hydrochloride and curcumin is 4-8:2-
4:1-3。
In a preferred embodiment, the weight proportion of the Bergenin, Berberine hydrochloride and curcumin is 5:3:
2。
The second aspect of the present invention is to provide described pharmaceutical composition and treats or prevents in disease of stomach drug in preparation
Using.
In one embodiment, the disease of stomach includes: chronic superficial gastritis, atrophic gastritis, gastric cancer
Preceding lesion.
It is used on the way in aforementioned pharmaceutical compositions, it can be by aforementioned pharmaceutical compositions system according to the animal state of an illness and agents area
For at suitable pharmaceutical preparation, to facilitate medication, for the present invention, the administration time and administration number of times of pharmaceutical composition are needed
Will be depending on the specific diagnostic result of the state of an illness, this is within the technical scope that those skilled in the art grasp.For example, will be to small
The therapeutic scheme of mouse is applied on the person, and all drugs can be by the drug to the effective dose of mouse to the effective dose of people
It converts, this is obvious for those of ordinary skill in the art.
The third aspect of the present invention provide include described pharmaceutical composition pharmaceutical preparation, by described pharmaceutical composition and
Pharmaceutically acceptable carrier composition.
In one embodiment, the pharmaceutical preparation is oral preparation, and the pharmaceutically acceptable carrier includes forming sediment
Powder, microcrystalline cellulose, sucrose, dextrin, lactose, glucose, magnesium stearate, stearic acid, sodium chloride, enuatrol, Abbas's sweet tea, pool
Luo Shamu, water, ethyl alcohol, hydroxypropyl methyl cellulose, sodium carboxymethylcellulose, sodium alginate, polyvinylpyrrolidone, bicarbonate
Sodium, citric acid, tartaric acid, hydroxypropyl cellulose, acacin, agar, alginic acid, cellulose ether, carboxymethyl crusta ester, poly- cream
Acid, polylactic-co-glycolic acid, polylactide-co-glycolide, polyglycolic acid, polyphosphazene, polybutylcyanoacrylate, in polyamide
It is one or more of.
In the present invention, Bergenin is also known as arolisic acid B, bergenin, is soluble in methanol, is slightly soluble in water, acetone and second
Alcohol is to extract from saxifragaceae plant isolated, and particular chemical formula is as follows:
It is existing research shows that Bergenin has the function of liver protection, anti-oxidant, anti-inflammatory, antibacterial, immunological regulation and hypoglycemic.
Berberine hydrochloride is also known as berberine, is present in many plants of Berberidaceae, is dissolved in water, is insoluble in benzene, ether and chlorine
Imitative, particular chemical formula is as follows:
Modern research shows that Berberine hydrochloride have decompression, heart tonifying, anti-arrhythmia, treat diabetes and its complication,
Antitumor, antimicrobial antiphlogistic isoreactivity.
Curcumin is the polyphenol compound extracted from the rhizome of some plants in Zingiber, Araeceae, is not dissolved in
Water and ether are dissolved in ethyl alcohol, propylene glycol, and particular chemical formula is as follows:
It is existing research shows that curcumin has reducing blood lipid, the protection of anti-oxidant, cardiovascular and cerebrovascular, anti-inflammatory isoreactivity.
The present invention is on the basis of discovery Bergenin has therapeutic effect to gastric precancerous lesion for the first time, in conjunction with active Chinese drug component
The literature survey of monomer is groped to obtain a kind of with the pharmaceutical composition obviously to act synergistically.Described pharmaceutical composition is viscous to stomach
Theca cell has dual regulation, and also has preferable therapeutic effect on rat gastric precancerous lesion model.
Specific embodiment
Also the present invention further can be understood by embodiment, wherein the embodiment illustrates some preparations or user
Method.It is to be appreciated, however, that these embodiments do not limit the present invention.The change of the invention of currently known or further exploitation
Change is considered within the scope of the invention described herein and claimed below.
In the present invention, the synergistic effect between drug refers to the biological effect after each group subassembly, and based on independent
It generates content required by given biological effect using expectation when single component to compare, above-mentioned composition can be by using
Lesser amount of component and obtain the biological effect, that is to say, that the activity of composition be apparently higher than single component superposition effect
It answers, i.e., produces synergistic effect between drug.
Influence of the example 1 drug composition to nitrosamine compound MNNG induction GES-1 malignant transformation of cells
The preparation of model group cell: human gastric epithelial cell line GES-1 is taken, with 1640 culture medium of RPMI containing 10%FBS
Subculture is carried out, presses 2 × 10 using the cell in logarithmic growth phase5A/ml concentration is inoculated in culture dish, 37 DEG C, 5%
CO22 × 10- is added in incubator after overnight incubation5The MNNG (N- methyl-N '-nitro-N nitrosoguanidine) of mol/L, is protected from light culture
Liquid is changed after 24 hours, culture after a week, collects residual cell, carries out culture biography with 1640 culture medium of RPMI containing 10%FBS
In generation, when reaching forth generation, collects cell as model group cell (MC cell).
GES-1 cell and MC cell are inoculated in 96 orifice plates with 4000/hole respectively, after being cultivated for 24 hours in incubator, respectively
Administration group adds relative medicine, and control group gives equivalent culture medium, and every group of each 4 hole is continued after cultivating 72h, measured using mtt assay
Group of cells activity calculates the inhibiting rate that each administration group grows GES-1 cell and MC cell, inhibiting rate=(cellular control unit
OD value-administration group cell OD value)/cellular control unit OD value × 100%.
As a result as follows:
1) each compound monomer influences the growth inhibition ratio of GES-1 cell
2) pharmaceutical composition of different weight proportion influences the growth inhibition ratio of GES-1 cell
Bergenin | Berberine hydrochloride | Curcumin | Growth inhibition ratio (%) | |
Group 1 | 8mg/L | 1mg/L | 1mg/L | -21.7±0.6 |
Group 2 | 6mg/L | 2mg/L | 2mg/L | -23.9±0.5 |
Group 3 | 5mg/L | 3mg/L | 2mg/L | -26.4±0.6 |
Group 4 | 3mg/L | 2mg/L | 5mg/L | -12.3±0.4 |
Group 5 | 2mg/L | 5mg/L | 3mg/L | 1.2±0.2 |
Group 6 | 1mg/L | 8mg/L | 1mg/L | 6.8±0.4 |
It can be seen from the results above that Bergenin and curcumin have a had significant proliferation effect to Gastric Mucosal Cells, and salt
Sour jamaicin has certain inhibiting effect in high dose.And pharmaceutical composition equally makees Gastric Mucosal Cells with had significant proliferation
With, but when content of berberine hydrochloride is excessively high, this proliferation function disappears.This prompt pharmaceutical composition is under approrpiate wts proportion
There is preferable protective effect to Gastric Mucosal Cells.
3) each compound monomer influences the growth inhibition ratio of MC cell
4) pharmaceutical composition of different weight proportion influences the growth inhibition ratio of MC cell
Bergenin | Berberine hydrochloride | Curcumin | Growth inhibition ratio (%) | |
Group 1 | 8mg/L | 1mg/L | 1mg/L | 18.6±0.4 |
Group 2 | 6mg/L | 2mg/L | 2mg/L | 38.1±0.6 |
Group 3 | 5mg/L | 3mg/L | 2mg/L | 61.4±0.5 |
Group 4 | 3mg/L | 2mg/L | 5mg/L | 50.9±0.4 |
Group 5 | 2mg/L | 5mg/L | 3mg/L | 56.3±0.6 |
Group 6 | 1mg/L | 8mg/L | 1mg/L | 46.2±0.4 |
MC cell is to induce GES-1 cell to obtain by MNNG, research shows that MC Chromosome aberration increases, bone
Frame microfilament is abnormal, and obtains soft-fractrue rock mass ability, but cannot be carcinogenic.It is therefore contemplated that MC cell is research gastric cancer
The preceding preferable cell model of lesion.It can be seen from the results above that pharmaceutical composition has preferable inhibit to MC cell Proliferation
Effect, in conjunction with GES-1 cell correlated results can be seen that pharmaceutical composition to gastric mucosal cell have dual regulation,
The reparation that normal gastric mucosa cell (GES-1) can not only be promoted can also inhibit the proliferation of MC cell after induced mutation, and
It is that 5:3:2 effect is best with Bergenin, Berberine hydrochloride and curcumin weight proportion.
5) influence of the drug to E-cadherin albumen and P-stat3 protein content in MC cell
Take above-mentioned MC cell inoculation in 24 orifice plates, being added drug culture according to the method described above, every group of two holes, culture
After 72h, supernatant is removed, group of cells is collected in digestion, cell pyrolysis liquid is added, after albumen is collected by centrifugation, using ELISA reagent
Box measures each group E-cadherin albumen and P-stat3 protein content respectively.
Concrete outcome is as follows:
* it is respectively represented compared with the control group with * *, P < 0.05 and 0.01 (Oneway-ANOVA)
The influence for the GES-1 cellular damage that 2 pharmaceutical composition of embodiment induces helicobacter pylori (Hp)
For gastric precancerous lesion, other than chemical substance induction is caused a disease, damage is in gastric cancer caused by helicobacter pylori
It is also an important pathogenic factor in preceding lesion.It is therefore desirable to the influences to pharmaceutical composition to helicobacter pylori induced damage
It is studied.
GES-1 cell is inoculated in 96 orifice plates with 4000/hole, after cultivating for 24 hours in incubator, Hp bacterium is added to each hole
Liquid (1 × 1011CFU/L) 12 μ l, blank group give equivalent culture solution, continue after cultivating 6h, and administration group adds related drugs solution,
Blank group and model group give equivalent culture solution, continue culture for 24 hours, carry out coherent detection 1) group of cells is measured using mtt assay
Activity calculates the activity of GES-1 cell, cell activity=administration group cell OD value/blank group cell OD value × 100%.2) it takes
Group of cells supernatant measures LDH and IL-1 β content, and LDH kit builds up Bioengineering Research Institute purchased from Nanjing, IL-1 β's
ELISA kit is purchased from U.S. abcam company.
Concrete outcome is as follows:
1) protective effect of the drug to Hp induced damage GES-1 cell
Bergenin | Berberine hydrochloride | Curcumin | Cell activity (%) | |
Model group | - | - | - | 45.9±5.1 |
Group 1 | 10mg/L | - | - | 55.4±5.2** |
Group 2 | - | 10mg/L | - | 69.3±4.9** |
Group 3 | - | - | 10mg/L | 48.6±4.3 |
Group 4 | 5mg/L | 3mg/L | 2mg/L | 91.9±4.7** |
Group 5 | 3mg/L | 2mg/L | 5mg/L | 77.1±5.2** |
* it is respectively represented compared with model group with * *, P < 0.05 and 0.01 (Oneway-ANOVA)
2) influence of the drug to Hp induced damage GES-1 cell LDH activity and IL-1 β level
* it is respectively represented compared with model group with * *, P < 0.05 and 0.01 (Oneway-ANOVA)
Influence of 3 pharmaceutical composition of embodiment to rat gastric precancerous lesion model
The SD rat of 200g or so is taken, it is random to be grouped, every group 10, the MNNG solution of 100mg/ml is freely drunk daily
(drinking bottle is protected from light processing) adds the mouse grain fodder of 0.03% ranitidine in feed, and stomach-filling gives 56 DEG C of 15% hot salt brine, often
Day is primary;Fasting in two days one day is fed, feed that morning stomach-filling gives 0.85% sodium deoxycholate solution (10ml/kg), prohibits
40% alcohol stomach-filling (10ml/kg) of that afternoon is eaten, blank group is without any processing;Modeling starts to be administered after 24 weeks, blank
Group and model group give physiological saline, and administration group presses 25mg/kg gastric infusion, and once a day, each group dosage regimen is as follows:
Bergenin | Berberine hydrochloride | Curcumin | |
Blank group | - | - | - |
Model group | - | - | - |
Group 1 | 25mg/kg | - | - |
Group 2 | - | 25mg/kg | - |
Group 3 | - | - | 25mg/kg |
Group 4 | 12.5mg/kg | 7.5mg/kg | 5mg/kg |
After being administered 12 weeks, puts to death rat and carry out Biological Detection.
Concrete outcome is as follows:
1) influence of the pharmaceutical composition to stomach lining pathomorphism
Stomach lining pathomorphism is observed under an optical microscope, is scored according to following standard, and slight: scaly epithelium increases
Thickness has certain atypia, and basement membrane is complete, and body of gland increased significantly, and lumen of gland expands, and gland cell core also increases, and atypia is unobvious;In
Degree: scaly epithelium thickens, and has certain atypia, and accidental nuclear fission, basement membrane is complete, and body of gland increased significantly, and lumen of gland expands, gland cell
Nuclear hyperchromatism has certain atypia, has no nuclear fission;Severe: scaly epithelium thickens, and has certain atypia, accidental nuclear fission, basement membrane
Completely, body of gland increased significantly, and lumen of gland expands, and gland cell nuclear hyperchromatism has certain atypia, it is seen that nuclear fission;As a result as follows:
Number of elements | Normally | Slightly | Moderate | Severe | |
Blank group | 10 | 10 | 0 | 0 | 0 |
Model group | 10 | 0 | 1 | 4 | 5 |
Group 1 | 10 | 3 | 1 | 3 | 3 |
Group 2 | 10 | 4 | 2 | 3 | 1 |
Group 3 | 10 | 2 | 3 | 2 | 3 |
Group 4 | 10 | 8 | 1 | 1 | 0 |
2) gastric analysis: after administration, 3 rats of every group of picking collect gastric juice, 3000rpm using pylorus ligature law
Centrifugation takes total acidity and pepsin activity in supernatant measurement gastric juice
3) mda content measures in gastric mucosa
After rat is put to death, gastral cavity is cut off along big curved side, filter paper blots mucosa surface moisture, and it is glutinous that slide scrapes paries posterior gastricus
Film, specific as follows using malonaldehyde (MDA) content in thiobarbituricacidα- method measurement mucous membrane:
The content of present invention merely illustrates claimed some specific embodiments, one of them or more skill
Documented technical characteristic can be combined with arbitrary one or more technical solutions in art scheme, these are combined and obtain
Technical solution also in the application protection scope, the technical solution just as obtained from these are combined is disclosed in the present invention
It is specifically recorded in content the same.
Claims (6)
1. the pharmaceutical composition of a kind of prevention or treatment gastric precancerous lesion comprising Bergenin, Berberine hydrochloride and curcumin,
The weight proportion of the Bergenin, Berberine hydrochloride and curcumin is 5:3:2.
2. pharmaceutical composition described in claim 1 treats or prevents the application in disease of stomach drug in preparation.
3. application according to claim 2, which is characterized in that the disease of stomach includes: chronic superficial gastritis, chronic
Atrophic gastritis, gastric precancerous lesion.
4. a kind of pharmaceutical preparation comprising claim 1 described pharmaceutical composition, by described pharmaceutical composition and pharmaceutically may be used
The carrier of receiving forms.
5. pharmaceutical preparation according to claim 4, which is characterized in that the pharmaceutical preparation is oral preparation.
6. pharmaceutical preparation according to claim 5, which is characterized in that the pharmaceutically acceptable carrier include starch,
Microcrystalline cellulose, sucrose, dextrin, lactose, glucose, magnesium stearate, stearic acid, sodium chloride, enuatrol, Abbas's sweet tea, pool Lip river
Sha Mu, water, ethyl alcohol, hydroxypropyl methyl cellulose, sodium carboxymethylcellulose, sodium alginate, polyvinylpyrrolidone, bicarbonate
Sodium, citric acid, tartaric acid, hydroxypropyl cellulose, acacin, agar, alginic acid, polylactic acid, polylactic-co-glycolic acid, poly- third are handed over
One or more of ester-glycolide, polyglycolic acid, polyphosphazene, polybutylcyanoacrylate, polyamide.
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