CN105902696B - Application of the viola mandshurica in the drug of preparation prevention and treatment II type virus infection of dengue fever - Google Patents

Application of the viola mandshurica in the drug of preparation prevention and treatment II type virus infection of dengue fever Download PDF

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Publication number
CN105902696B
CN105902696B CN201610508598.4A CN201610508598A CN105902696B CN 105902696 B CN105902696 B CN 105902696B CN 201610508598 A CN201610508598 A CN 201610508598A CN 105902696 B CN105902696 B CN 105902696B
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drug
viola mandshurica
water extract
treatment
virus infection
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CN105902696A (en
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李耿
赖小平
吴建国
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Guangzhou University of Chinese Medicine
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Guangzhou University of Chinese Medicine
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/86—Violaceae (Violet family)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Alternative & Traditional Medicine (AREA)
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  • Botany (AREA)
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  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Medicinal Preparation (AREA)

Abstract

The present invention relates to application of the viola mandshurica in the drug of preparation prevention and treatment II type virus infection of dengue fever.Viola mandshurica water extract in application of the present invention has obvious inhibiting effect to dengue type-2 virus, can be used for preparing the drug of prevention and treatment II type virus infection of dengue fever.The drug of prevention and treatment II type virus infection of dengue fever of the present invention is clinically acceptable solid or liquid oral medicine made of viola mandshurica water extract and the medically acceptable auxiliary material of conventional amount used.

Description

Application of the viola mandshurica in the drug of preparation prevention and treatment II type virus infection of dengue fever
Technical field
The present invention relates to containing the pharmaceutical preparation for not determining structure from plant, and in particular to Violaceae Viola is pale reddish brownly New application of the fourth plant in pharmacy.
Background technique
Dengue virus belongs to flaviviridae, Flavivirus in classification.Including 4 different serotypes, i.e., I, II, III, IV type Dengue virus.Dengue fever is the acute infectious disease as caused by dengue virus, is mainly propagated by Aedes aegypti and aedes albopictus, is passed The approach of broadcasting is infected after yellow-fever mosquito bites the blood of viremia virusemia patient.Clinical manifestation is mainly high fever, headache, muscle and joint Bitterly, fash, enlargement of lymph nodes and oligoleukocythemia etc., serious person may occur in which bleeding or shock, or even dead.The disease Major Epidemic In subtropical and tropical zones, there are case in existing more than the 100 a countries and regions in the whole world at present, have about 25~3,000,000,000 people to be in In the danger of dengue virus infection.The disease has seriously affected the health and socio-economic development of people.Due to current It there is no dengue vaccine, also lack corresponding antiviral drugs, how to cope with this emerging infectious disease and have become medicine sector urgently The key subjects that need to be coped with.
Viola mandshurica (Viola yedoensis) is Violaceae (Violaceae) Viola (Viola L.) perennial grass This plant also known as light calyx violet, viola japonica, Bunge Corydalis Herb and Herba Violae etc..Taste bitter, acrid, cold.Distributed in heart and lung channels.With heat-clearing solution Poison, cool blood detumescence, the effect of clearing heat and promoting diuresis cure mainly furunculosis, carbuncle swells, scrofula, jaundice, dysentery, diarrhea, hot eyes, sore throat, and poisonous snake stings Wound.Modern pharmacology research finds that it has preferable antiviral, anti-inflammatory, antibacterial and anti-tumor activity.Clinically it is chiefly used in controlling Treat furuncle swelling toxin, urinary tract infections, cellulitis, mammitis, pharyngitis, dysentery, jaundice, red eye, swell pain etc..Document report at present In there are no viola mandshurica anti-dengue virus active report and its application in terms of anti-dengue virus occur.
Summary of the invention
The new application that technical problem to be solved by the invention is to provide viola mandshuricas in pharmacy.
Above-mentioned new application is specifically application of the viola mandshurica in the drug of preparation prevention and treatment II type virus infection of dengue fever.
In above-mentioned application, the drug is the medically acceptable auxiliary material system of viola mandshurica water extract and conventional amount used At clinically acceptable solid or liquid oral medicine.
In above-mentioned application, the solid orally ingestible is specifically clinically acceptable granule, capsule or tablet.
In above-mentioned application, the liquid oral medicine is specifically clinically acceptable oral solution.
In above-mentioned application, the viola mandshurica water extract can be used method commonly used in the art and extract to obtain.The present invention The method that people recommends is as described below:
It is that solvent uses conventional refluxing extraction to obtain that water extract of the present invention, which is with water, wherein the present inventor recommends Extracting method difference it is as described below.
Viola mandshurica medicinal material is taken, 10 times of water are added, refluxing extraction 30 minutes, let cool, filters, takes filtrate, is concentrated, it is dry, Up to viola mandshurica water extract.
The preparation method of solid orally ingestible of the present invention is the customary preparation methods of defined in pharmacopeia, i.e., will be purple Flower Chinese violet water extract is mixed to prepare with the common auxiliary material of solid orally ingestible by clinically acceptable proportion.
The preparation method of liquid oral medicine of the present invention is the customary preparation methods of defined in pharmacopeia, i.e., will be purple Flower Chinese violet water extract is mixed with the common auxiliary material of liquid oral medicine by clinically acceptable proportion after being diluted with water, and is stirred evenly, is filtered Cross, it is filling, sterilizing to get.
Solid orally ingestible or liquid oral medicine of the present invention have the function of anti-II type dengue fever virus, can use In prevention and treatment dengue infection.
Drug safety and beneficial effect of the invention are further illustrated below by way of experiment.
The present invention is to be made using cytopathic-effect inhibition assay research viola mandshurica water extract to the inhibition of dengue fever II type virus With observation toxic effect of the viola mandshurica water extract to BHK-21 cell and the inhibiting effect to dengue fever II type virus utilize CCK-8 detection reagent measures the toxic concentration (CC of half of test medicine50) it is 35.21, half maximal effect concentration (EC50) be 12.92, selecting index SI is 2.73, efficiently for low toxicity.It is described that specific experimental method is as follows.
1. experimental material
1.1 cell strain cream hamster kidney cells (BHK-21), quoted from Wuhan institute of viruses, T11 generation.This room freezes conservation.
1.2 experimental groups and its corresponding test medicine
Given the test agent: the viola mandshurica water extract for taking following embodiments 1 to prepare is dissolved in water, and is made and is equivalent to every milliliter and contains The medical fluid of crude drug 100mg.
1.3 Strain dengue type-2 virus are quoted from Wuhan institute of viruses
1.3 reagents and instrument newborn calf serum and DMEM culture medium (Gibco company)；DMSO (Guangzhou Kang Yang chemical industry Co., Ltd)；Trypsase (DIFCO company, the U.S.；Shanghai Sheng Gong bio-engineering corporation agency).Olympus PM-6 is inverted aobvious Micro mirror (Japanese OLYMPUS company)；；E-52AA Rotary Evaporators (Shanghai Yarong Biochemical Instrument Plant)；BP221S electronic analysis day Flat (Shanghai precision instrument table Co., Ltd).
2. method
The measurement of 2.1 viola mandshurica water extract toxic concentrations on BHK-21 cell
It is 0.5 × 10 by density5Cell/ml BHK-21 cell inoculation is in 96 porocyte culture plates, every 100 μ L of hole.It is raw Long cell for 24 hours is adherent rear spare in flakes (80-90%).By drug with cell maintenance culture solution (contain 2% serum) from continuous 2 times of ladders Degree 6 gradients of dilution, 100 μ L of institute's preparating liquid is added in every hole, if normal cell controls, every hole is added 100 μ L cells and maintains training Nutrient solution.If solvent control group, in 37 DEG C of 5%CO2It is cultivated in incubator.It is added after dosing culture 48h and contains 100 μ L10%CCK- The culture medium of 8 reagents, in 37 DEG C of incubation 1h.It is calculated according to following method:
Absorbance is detected with 450nm, drug indicates the toxicity of cell with the activity of cell, calculation formula:
Cell viability (%)=(medicine group-blank control)/(cell controls-blank control) × 100%.
The measurement of 2.2 viral suppressions
Selection grows vigorous, layered good cell, and pancreatin digestion is added in tipping growth-promoting media, makes cell with growth-promoting media dispersion Suspension concentration is 105A/hole.Cell suspension is added in 96 orifice plates with pipettor, every hole 100uL.Culture plate is placed in culture It is cultivated 12~18 hours in case, after cell grows up to single layer in 96 orifice plates, sucks culture solution, the difference of 100ul is added in every hole The drug of diluted concentration and the 100TCID of 100uL50Virus liquid, 2 multiple holes are arranged in each drug concentration, while blank pair is arranged It according to group, normal cell controls group, virus control group and positive controls, is placed in incubator and is incubated for 1h, suck drug containing culture solution And virus liquid.Maintaining liquid is sucked in virus control group 75%-100% lesion, 100 μ L 10%CCK-8 is added, in microplate reader 450nm measures absorbance, calculates the viral suppression of drug.
Viral suppression=(test group mean OD value-virus control group mean OD value)/(cell controls group mean OD value- Virus control group mean OD value) × 100%
3. result
3.1 drug toxicity results such as table 1.
Table 1
Measurement result such as table 2 of 3.2 drugs to viral suppression.
Table 2
It is above-mentioned the experimental results showed that water extract of the present invention has obvious inhibiting effect to dengue type-2 virus.Select index Greater than 2, it can be used for preparing the treatment and prevention drug of dengue fever.
Specific embodiment
Example 1:
Dry viola mandshurica herb 10kg is taken, adds 100L distilled water immersion 30 minutes, decocts 30 minutes, filters while hot, It is concentrated into the thick medicinal extract of 1.05~1.1 specific gravity, it is dry, obtain viola mandshurica water extract.
Example 2:(granule)
The auxiliary materials such as sucrose, dextrin are added in the water extract 300g of Example 1, are placed in fluid bed granulator and mix, pelletize, It is dry, packing, every bag of 9g.This product is brown green coloured particles, slightly sweet, bitter.Other projects should meet Pharmacopoeia of People's Republic of China Pertinent regulations under version granule item in 2010.
Example 3:(capsule)
The auxiliary materials such as starch are added in the water extract 300g of Example 1, are placed in fluid bed granulator and mix, pelletize, dry. Gained particle is fitted into No. 1 hard gelatin capsule, every 0.3g, dispenses 100.This product is capsule, and content is brown-green Particle or powder, mildly bitter flavor, for being administered orally.Other projects should meet Pharmacopoeia of People's Republic of China version capsule in 2010 Pertinent regulations under.
Example 4:(tablet)
The auxiliary materials such as starch are added in the water extract 100g of Example 1, are placed in fluid bed granulator and mix, pelletize, after dry Appropriate magnesium stearate tabletting is added, every 0.2g dispenses 500.This product is brown-green tablet, mildly bitter flavor, for being administered orally.Its Its project should meet the pertinent regulations under Pharmacopoeia of People's Republic of China tablet item in 2010.
Example 5:(oral solution)
The water extract 50g of Example 1, adds appropriate amount of water to dissolve, and filtration is added simple syrup and appropriate sodium benzoate, adds water It to 1000ml, stirs evenly, stands, filtration, encapsulating, sterilizing, every 10ml.This product is brown supernatant liquid, and sweet, slight bitter is for oral It uses.Other projects should meet the pertinent regulations under Pharmacopoeia of People's Republic of China oral solution item in 2010.

Claims (3)

1. viola mandshurica water extract answering in the drug of preparation prevention and treatment II type virus infection of dengue fever as sole active agent With wherein the viola mandshurica water extract is prepared by following methods:
Viola mandshurica is taken, 10 times of water are added, refluxing extraction 30 minutes, let cool, filters, takes filtrate, is concentrated, it is dry to get pale reddish brown Chinese violet water extract.
2. application according to claim 1, characterized in that the drug is by viola mandshurica water extract and conventional amount used Medically acceptable auxiliary material made of clinically acceptable solid or liquid oral medicine.
3. application according to claim 2, characterized in that the solid or liquid oral medicine is granule, capsule Agent, tablet or oral solution.
CN201610508598.4A 2016-06-29 2016-06-29 Application of the viola mandshurica in the drug of preparation prevention and treatment II type virus infection of dengue fever Active CN105902696B (en)

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