CN109953994A - Purposes of the Bolbostemma paniculatum glucoside A as treatment herpes simplex infections disease - Google Patents
Purposes of the Bolbostemma paniculatum glucoside A as treatment herpes simplex infections disease Download PDFInfo
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Abstract
Purposes the present invention relates to Bolbostemma paniculatum glucoside A (glycosides first) as treatment herpes simplex virus I-type (HSV-I) and II type (HSV-II) infectious disease.The rhizoma bolbostemmae is the dry tuber of cucurbitaceous plant Bolbostemma paniculatum [Bolbostemma paniculatum (maxim) Franquet (Cucurbitaceae)], is a kind of traditional Chinese medicine.The glycosides first isolated from the rhizoma bolbostemmae is a kind of pentacyclic triterpene saponin with unique texture, relative molecular weight 1318Da, molecular formula C63H98O29, purity >=95%.Glycosides first has the activity for significantly inhibiting HSV-I and HSV-II virus multiplication；Glycosides first has the activity for tri- gene duplications of UL27, UL52 and UL54 for significantly inhibiting two herpes simplex virus strains of HSV-I/F and HSV-II.The glycosides first of nontoxic high concentration also has inhibitory activity to the proliferation of HSV-I/106 persister.Observation indicate that glycosides first has significant curative effect to skin bleb caused by HSV-I and HSV-II.Glycosides first is to the nonirritant reaction of rabbit skin, to cavy without active cutaneous anaphylaxis.Stability test shows that glycosides first sets preservation at dry, and the shelf-life is 1.0 years.
Description
Background of invention
Herpes simplex virus (herpes simplex virus, HSV) can be divided into I type and II type (HSV-I and HSV-
II).People is the unique natural reservoir (of bird flu viruses) of HSV.I type herpe simplex disease mainly passes through respiratory tract, skin and mucous membrane close contact and passes
It broadcasts, infects waist with the mucocutaneous and organ of upper bit, such as cause the inflammation of lip mucosa, vestibulum nasi, eye conjunctiva, bottleneck throat
And bleb.The bleb occurred around mouth and mouth, 99% is as caused by I type herpesvirus infection.It can clinically be divided into again primary
Type and recurrent herpes bleb.Common person for example skin bleb, herpetic gingivostomatitis, herpes ophthalmicus (acute conjunctivokeratitis,
Ulcer of the cornea), eczema sample bleb, herpetic paronychia, severe one can develop as general herpes simplex, herpetic rectitis, urgency
Property encephalitis, acute cerebral meningitis and ramitis etc..
HSV-II type mainly causes genitals and waist following skin bleb.If pregnant woman suffers from primary herpesviral sense
Dye, virus are possible to form congenital infection by placental infection fetus during viremia virusemia.If pregnant woman's birth canal sense
It contaminates herpesviral (primary or recurrent infection), then virus can infect newborn in birth process and cause infection of newborn.Nothing
By being congenital infection or infection of newborn, prognosis is all poor.In addition, the disease incidence of herpes simplex virus is only secondary in venereal disease
In stranguria syndrome and syphilis, the third position of venereal disease disease incidence is occupied.
Neutrality antibody is generated in vivo, but cannot prevent from recurring within 4~5 days after primary infection.Primary patient can have fever,
Body temperature may be up to 39 DEG C, the course of disease 7~9 days；Bleb secondary bacterial infection is then in warts sample or eczema sample, the course of disease extend and more after
Skin stays scar.Recurrent herpes simplex can have cellular immunity deficiency.HSV-I type is hidden in gasserian ganglion more, and HSV-
II type then more hides in waist sacrum dorsal ganglion, when body over fatigue, fever, gastrointestinal dysfunction, menstruation and the immune suppression of application
When preparation, latent HSV can be activated and fall ill.
To various mucocutaneous diseases caused by HSV-I and HSV-II, now clinically commonly use acyclovir, Valaciclovir,
Ganciclovir, interferon, transfer factor and interleukins whole body and/or local treatment.These drugs have certain curative effect,
Normal recurrent exerbation after healing.
The rhizoma bolbostemmae [Bolbostemma paniculatum (Maxim.) Franquet (Cucurbitaceae)] is a kind of
Traditional Chinese medicine, is loaded in the compiled supplementary Amplifications of the Compendium of Materia Medica of Qing Dynasty Zhao Xueminshi, and medicinal effects are its bulb.It is controlled in ancient times with it
The various illness such as mammary cancer, acute mastitis, the proliferation of mammary gland and scrofula are treated, clinically show apparent dissipating bind poison, subduing inflammation effect.In
It is medical it treat cancer of the esophagus, gastric cancer etc., eliminate obstruction, reduce lump, extending life significant effect.But because the rhizoma bolbostemmae is raw
Medicine decoction often causes the symptoms of digestive tract such as Nausea and vomiting, and some patients are not easy-tolerated.
Bolbostemma paniculatum glucoside A (calling glycosides first in the following text) is the monomer for separating, being purified into from rhizoma bolbostemmae bulb, purity >=95%,
Chemical structure has determined that (Fig. 1), relative molecular weight 1318Da.Glycosides first has unique pentacyclic triterpene saponin structure, sour water
Bei e aglycon (bayogenin) and four kinds of sugar: glucose, rhamnose, xylose and arabinose, molar ratio 1 are generated after solution
∶1∶1∶2。
Brief summary of the invention
Purposes the present invention relates to glycosides first as treatment HSV-I and HSV-II infectious disease.Experimental fact is provided below
(including method and result) has as glycosides first inhibits HSV-I and HSV-II proliferation and its active evidence of gene duplication；It enumerates
Preliminary observation case can be used as the evidence for the treatment of HSV-I and HSV-II infectious disease candidate medicinal usage as glycosides first；Glycosides is provided
First the skin irritation test of experimental rabbit, glycosides first are given the test of the active skin anaphylactic test of cavy as a result, as glycosides
First can be used as treating the evidence of HSV-I and HSV-II infectious disease candidate drug external application safety；Glycosides first stability test knot is provided
Fruit can keep stable evidence as drug as glycosides first.
One, anti-HSV-II and the HSV-II activity of glycosides first
One) material
1, positive reference substance: acyclovir (ACV): purity > 98%, molecular weight: 225.2, it is purchased from Med Chem
Express, article No.: 59277-89-3.
2, glycosides first: white powder, purity >=95%.It is appropriate that precision weighs sample, is added after tri-distilled water is completely dissolved and sterilizes
Processing, dispenses 20 μ L/ branch, and -20 DEG C of conditions save backup.
3, cell line and its culture: African green monkey kidney cell (vero cell), with DMEM culture medium (10% tire ox of addition
Serum, 100U/mL penicillin and 0.1mg/mL streptomysin) in 5%CO2, the interior culture of 37 DEG C of incubators.
4, Strain
4.1 F plants of herpes simplex virus type 1 (HSV-I/F), Wuhan Virology Institute,Chinan academy of Sciences's present.
4.2 herpes simplex virus type 2 Strain (HSV-II), Wuhan Virology Institute,Chinan academy of Sciences's present.
4.3 herpes simplex virus type 1 acyclovir multidrug resistant disease strains (HSV-I/106), Chinese Academy of Sciences's Guangzhou biology doctor
Medicine and health research institute present.
Two) experimental method
1, cytotoxicity of the glycosides first to African green monkey kidney cell (vero cell)
The initial concentration of glycosides first toxicity test is 100 μ g/mL, successively 7 concentration gradients of doubling dilution.With it is inoculated
Vero cell is incubated with, and uses MTT method detection compound to the toxic effect of vero cell for 24 hours, after 48h, 72h.
2, the virulence of herpes simplex virus strain
2.1 virus amplification
Virus stain is seeded to the vero cell of logarithmic growth phase, continues culture to cell and complete lesion occurs. -80
, collect virus multiplication liquid, packing to viral cryopreservation tube, 200 μ L/ pipe is placed -80 DEG C and saved backup DEG C multigelation for 3 times.
2.2 virus virulence
2.2.1 viral median tissue (cell) culture infective dose (TCID50)
With maintaining liquid by virus stock solution used carry out 10 times be serially diluted after, inoculation is seeded to the cells of 96 orifice plates, every hole in advance
100μL.Cell controls group adds 100 μ L cell maintenance mediums.37 DEG C, 5%CO2Culture in incubator.Appearance is observed and recorded daily
The hole count of cytopathy (cytopathic effect, CPE), cell is disease-free to be become-, lesion 0~25% be+, lesion 25~
50% is ++, lesion 50~75% is +++, lesion 75~100% is ++++.When cytopathy not developing deeply, Reed- is used
Muench formula calculates 50% tissue cytopathogenic dose (LgTCID50)。
LgTCID50=- [50% multiple of CPE >]+[(percentage -50% of > 50%)/(percentage < of > 50%
50% percentage)] × -1 (index that 50% multiple of CPE > is dilution).
2.2.2 plaque test titre
In cell inoculation to 24 orifice plates, every hole 1mL cell suspension.Viral dilution is 50,000,100,000,200,000,400,000,80
Ten thousand, the viral dilution of every 100 μ L of hole, 4 multiple holes of each concentration.Cell covering liquid 1mL is added after 2 h of viruses adsorption.According to
The formation size of scab, is dyed.Covering liquid is outwelled, the fixed 30min of formalin of 500 μ L is added.Formaldehyde is outwelled to fix
500 μ L of crystal violet application liquid is added in liquid, dyes 30min.Tap water rinses, and plaque is presented, and counts.
3, the antiviral activity (CPE record) of glycosides first
Glycosides first detects the inhibitory effect of virus multiplication by cytopathy (CPE) caused by observation virus.From right
Direct inactivation, the treatment and prevention of virus act on the antiviral activity of three aspect detection glycosides first.Wherein direct deactivation into
The detection of bis- incubation time points of row 2h, 4h, prevention experiment carry out the detection of tri- incubation time points of 6h, 12h, 18h.
4, virus plaque, which is formed, inhibits
Virus plaque, which is formed, inhibits testing inspection glycosides first to the antiviral activity of type strain.
5, real-time quantitative PCR (Real Time-PCR)
Fluorophor is added in PCR reaction system, record fluorescence signal accumulates the entire PCR process of real-time monitoring.It utilizes
Standard curve carries out quantitative analysis to unknown template.Glycosides first or ACV are added in the PCR reaction system that fluorophor is added,
The influence that glycosides first or ACV replicate HSV-I/F plants and HSV-II viral gene UL27, UL52, UL54 can be monitored.
Three) experimental result
1, cytotoxicity of the glycosides first to African green monkey kidney cell (vero cell)
CC of the glycosides first to the toxicity of vero cell50For 28.73 μ g/mL, CC95For 0.66 μ g/mL, CC0For 0.55 μ g/mL
(table 1, Fig. 2).
Toxicity of 1 Bolbostemma paniculatum glucoside A of table to vero cell
2, virus virulence
Virus virulence measurement result is shown in Table 2.
The virulence of 2 virus stain of table
Note: PFU, plaque forming unit (plaque forming unit).
3, the antiviral activity of glycosides first
Direct deactivation of the 3.1 glycosides first to HSV-I and HSV-II standard strain
Glycosides first has significant direct deactivation to HSV-I and HSV-II virus.Directly it is incubated for 4h, minimum in experiment
The glycosides first of 0.0244 μ g/m of concentration still can completely or significantly inhibit cell that infection lesion occurs.
The inhibitory effect of 2h is directly inactivated lower than directly inactivation 4h.CPE the results are shown in Table 3.
Direct deactivation of 3 Bolbostemma paniculatum glucoside A of table to HSV-I/F and HSV-II
Note: CPE observation :-indicate that cell-free lesion generates；+ indicate that≤20% cell finds lesion；++ indicate 20%
~50% cytopathy；+++ indicate 50%~75% cytopathy；++++indicate 75% or more cytopathy.
Therapeutic effect that 3.2 glycosides first infect HSV-I and HSV-II standard strain,,,,,,
Glycosides first significantly inhibit HSV-I/F and HSV-II strain cause cytopathy (cell rounding after virus infected cell,
Swelling and fall off), and the inhibitory effect of HSV-I/F is apparently higher than to HSV-II (table 4).
Therapeutic effect of 4 Bolbostemma paniculatum glucoside A of table to HSV-I and HSV-II type strain
Note: CPE observation :-indicate that cell-free lesion generates；+ indicate that≤20% cell finds lesion；++ indicate 20%
~50% cytopathy；+++ indicate 50%~75% cytopathy；++++indicate 75% or more cytopathy.
Prevention effect of the 3.3 glycosides first to HSV-I and HSV-II type strain
Glycosides first with cell incubation 6h, 12h, 18h, inoculates virus in advance.CPE testing result shows that glycosides first is to HSV-I/
F plants and activity of the HSV-II strain without significant inhibition cytopathy.It the results are shown in Table 5 and table 6.
5 Bolbostemma paniculatum glucoside A of table is to HSV-I/F plants of prevention effect
Prevention effect of 6 Bolbostemma paniculatum glucoside A of table to HSV-II type strain
Note: CPE observation :-indicate that cell-free lesion generates；+ indicate that≤20% cell finds lesion；++ indicate 20%
~50% cytopathy；
+++ indicate 50%~75% cytopathy；++++indicate 75% or more cytopathy.
4, glycosides first inhibits HSV virus plaque to be formed
Plaque reduction assay of the 4.1 glycosides first to HSV-I/F and HSV-II type strain
Plaque test subtrahend experimental result shows that glycosides first has the proliferation of HSV-I/F plants and HSV-II strain significant
Inhibitory activity, glycosides first respectively reach 69.86% and 56.16% to the plaque test maximal percentage inhibition of HSV-I/F and HSV-II,
As a result see Fig. 3.The glycosides first (0.39 μ g/mL and 0.78 μ g/mL) of nontoxic high concentration also has inhibition to HSV-I/106 persister
Activity.
5, the influence that glycosides first replicates HSV-I/F plants and HSV-II viral gene
Testing result shows, glycosides first can significantly inhibit HSV-I/F and HSV-II blister sore strain UL27, UL52 and
The duplication of tri- genes of UL54.As a result see Fig. 4 and Fig. 5.
Four) experiment conclusion
1, glycosides first has the activity for inhibiting herpesviral HSV-I/F and HSV-II to be proliferated significantly；
2, glycosides first significantly inhibits answering for tri- genes of UL27, UL52 and UL54 of herpesviral HSV-I/F and HSV-II
System.
3, the higher glycosides first of nontoxic but concentration (0.39 μ g/mL and 0.78 μ g/mL) has suppression to HSV-I/106 persister
System activity.
Two, preliminary observation case
This group observation case is hair in the skin bleb of skin and mucosa intersection.Bleb betides antelabium, bicker, performance
For the small protuberance of local skin, scleroma is formed.Have itch, scorching hot or shouting pain etc. initial symptoms person 6, there is congested, blush person 8
Example, occurs blister person 10, totally 24, divides 2 groups, each 12.The glycosides first aqueous solution or glycosides of 0.5mg/mL are dipped with cotton balls respectively
First and second alcoholic solutions, are applied to part, and every 2~3h is smeared 1 time.Treatment is smeared through glycosides first aqueous solution on the 1st or the first and second alcoholic solution of glycosides,
The symptoms such as the itching, is scorching hot of initial stage case, shouting pain disappear；It was treated through 2 days, congested, blush, the signs such as scleroma all subside；Through
It treats within 2nd~3, blister disappears.The therapeutic effect no significant difference of glycosides first aqueous solution, the first and second alcoholic solution of glycosides.
Three, the safety testing of glycosides first
1,50% cytotoxic concentration (CC of glycosides first50) measurement
Experiment uses the strain of T- Lymphoblastoid (MT-2 cell).Mtt assay testing result shows, the concentration of glycosides first
When dropping to 40 μ g/mL from 80 μ g/mL, cytotoxicity is reduced rapidly.Glycosides first is to 50% cytotoxic concentration of MT-2 cell
59.8μg/mL。
2, the acute toxicity test of glycosides first
Experiment does acute toxicity testing using adult ICR healthy mice, half male and half female, conventional method.The results show that glycosides
Median lethal dose (the LD of first intramuscular injection50) it is 39.3mg/kg；The LD of intraperitoneal injection50For 20.7mg/kg；The LD of stomach-filling50For
308.8mg/kg。
3, skin irritation test of the glycosides first to experimental rabbit
It entrusts Hunan Province's Experimental Animal Center (Drug Safety Evaluation Center of Hunan Province), implements glycosides first to experimental rabbit
Skin irritation test.According to national GLP code requirement, the experimental rabbit that quarantine is qualified, processing is lost hair or feathers at back is taken, male and female are each
Half, intact skin group and damaged skin group are divided into using section random device by gender weight, every group 4.Intact skin group exists
Every experimental rabbit left dorsal even spread glycosides first 75% ethanol solution (10mg/mL) 0.5mL on plucked intact skin,
75% ethyl alcohol 0.5mL is coated on plucked intact skin on the right side of every experiment rabbit back；Damaged skin group is tested at every
Even spread glycosides first 75% ethanol solution (10mg/mL) 0.5mL on plucked damaged skin on the left of rabbit back, in every reality
Test rabbit back right side plucked 75% ethyl alcohol 0.5mL of damaged skin even spread.With glassine paper, two layers of gauze and stingless
Swash immobilization with adhesive tape, the medicament contact skin time is 4h, wrappage is then removed, and clean medicine-feeding part with warm water, daily by upper
Method is administered once, and continuous 5 days in same regional administration.The administration phase after removing drug 1h and again coating before observe respectively
And situations such as recording every experimental rabbit medicine-feeding part erythema, oedema, pigmentation, blutpunkte, pachylosis or epidermatic atrophy,
And it occurs and regression time, and scores erythema and oedema.After last coating, 30 after removing drug~
60min, 24,48 and 72h, situations such as visually observing and record coating part whether there is or not erythema and oedema.
The results show that side is administered in glycosides first intact skin administration side, damaged skin, 75% ethyl alcohol intact skin control sides are broken
Skin control sides are damaged, coating localized skin surface visually observes and is showed no erythema, oedema, erosion, necrosis, and subcutaneous tissue is also not
The variations such as the swollen, necrosis of water breakthrough.
Conclusion: under this experimental condition, ((10mg/mL) is administered once daily (0.5mL/ is only) to glycosides first, and same position connects
Experiment rabbit skin is reacted in continuous administration 5 days without obvious irritation.
4, glycosides first gives the active skin anaphylactic test of cavy
It entrusts Hunan Province's Experimental Animal Center (Drug Safety Evaluation Center of Hunan Province), studies glycosides first to cavy
Active skin anaphylactic test.According to national GLP code requirement, the albino guinea-pig of quarantine qualification, 300~400 g of weight is chosen, it is female
It is male fifty-fifty, it is divided into negative control group, positive controls, glycosides first group, every group 6.Experiment the previous day carries out depilation processing.?
0,7,14 days, by 75% ethanol solution (10mg/ of 75% ethyl alcohol, 1%2,4- dinitrofluorobenzene, 70% ethanol solution and glycosides first
ML), be coated in back hair removal section on the left of corresponding animal respectively, 0.5mL/ only, glassine paper and two layers of gauzes covering, then with stingless
Swash property adhesive plaster and bandage is fixed.Every animal sub-cage rearing removes drug with warm water after 6h.Last gives tested material sensitization
14 days afterwards, in excitation contact phase, by 75% ethyl alcohol, 0.1%2,4- dinitrofluorobenzene, 70% ethanol solution, 75% second of glycosides first
Alcoholic solution (10mg/mL), 0.5mL/ only, are applied to hair removal section on the right side of corresponding animal, glassine paper and two layers of gauze covering respectively,
It is fixed again with nonirritant adhesive plaster and bandage.Remove drug with warm water after 6h, i.e., whether comparable, observation glycosides first causes
Cavy active cutaneous anaphylaxis.Then in for 24 hours, 48h, 72h observe again.
The results show that the positive controls of coating 1%2,4- dinitrofluorobenzene, allergic reaction sensitivity response rate is 100%
(6h), 100% (for 24 hours), 50% (48h) and 16.7% (72h), hypersensitive evaluation are respectively extreme sensitization (6h), pole
Spend sensitization (for 24 hours), moderate sensitization (48h) and slight sensitization (72h)；It is coated with the negative control group of 75% ethyl alcohol, allergy
React 6h, for 24 hours, 48h and 72h sensitivity response rate be 0, hypersensitive evaluation be no sensitization；It is coated with glycosides first group, mistake
Quick reaction 6h, for 24 hours, 48h and 72h sensitivity response rate be 0, hypersensitive evaluation be no sensitization.
Conclusion: under this experimental condition, glycosides first (10mg/mL) has no obvious active cutaneous anaphylaxis to cavy.
5, acute eye irritation/corrosion test
The preparation of 5.1 glycosides first solution: with normal saline at the glycosides first aqueous solution of 1mg/mL, 3mg/mL, 5mg/mL,
Filtration sterilization.
5.2 animals: experimental rabbit, regular grade, 1.9~2.2kg, male and female have concurrently.Test before for 24 hours to the eyes of every rabbit into
Row checks.Reject the animal for having Eye irritation symptom, corneal defect and conjunctival damage.
5.3 animal packets: glycosides first solution 1mg/mL, 3mg/mL, 5mg/mL group, totally 3 groups, every group of 3 animals.
5.4 medications: using androgynous left and right self-contrast method.The left eye of every rabbit of each group instills 0.1mL physiology salt
Water；Glycosides first (1mg/mL) organizes the glycosides first solution 0.1mL that right eye instills 1mg/mL；Glycosides first (3 mg/mL) organizes right eye and instills 3mg/mL
Glycosides first solution 0.1mL；Glycosides first (5mg/mL) organizes the glycosides first solution 0.1mL that right eye instills 5mg/mL.Then it gently sleeps eyelid about 10
Second.
5.5 check: upon administration 1,2,4,24,48,72h checks eye.
5.6 results: instilling the right eye of 1mg/mL, 3mg/mL, 5mg/mL glycosides first solution 0.1mL and instills 0.1mL physiology salt
The left eye of water is the same, and cornea, iris and conjunctiva do not observe anomalous variation.
5.7 conclusions: under this experimental condition, the disposable eye drip of glycosides first solution (1mg/mL, 3mg/mL, 5mg/mL) 0.1mL
Obvious irritation, which is reacted, to be had no to experiment lagophthalmos.
Four, the stability test of glycosides first
1, influence factor is tested
1.1 hot test
Glycosides first 30mg, 40mg, 50mg are taken, is set in sealing clean container, is placed 10 days under the conditions of 60 DEG C, in 0,5,10
Its sampling, by the purity of pharmacopeia chromatography detection glycosides first.The results show that the purity of glycosides first is without significant changes.
1.2 high humidity test
Glycosides first 1.01g, 1.08g, 0.92g are taken, constant humidity (KNO is set3Saturated solution, RH92.5%) in chromatography cylinder, at 25 DEG C
Under the conditions of it is closed place 10 days, in sample detection moisture absorption in 0,5,10 day increase weight.It is inhaled as a result, it has been found that glycosides first is average at the 5th, 10 day
Wet weight gain reaches 5.1 ± 0.2%.It can be seen that glycosides first have it is certain draw it is moist.
Take glycosides first 0.85g, 0.94g, 0.96g, set in constant humidity (NaCl saturated solution, RH75%) chromatography cylinder, 25 DEG C~
It places 10 days under the conditions of 32 DEG C, increases weight in sample detection moisture absorption in 0,5,10 day, and by the purity of pharmacopeia chromatography detection glycosides first.
As a result, it has been found that average moisture absorption weight gain below 5% (the moisture absorption weight gain 3.8 ± 0.6%) at the 5th, 10 day of glycosides first, purity is without aobvious
Write variation.
The test of 1.3 strong illuminations
Method: taking glycosides first 20mg, 30mg, 40mg, set in the lighting box equipped with fluorescent lamp, the temperature in test in lighting box
It spends consistent with room temperature.It places 10 days, was sampled at 0,5,10 day, by pharmacopeia chromatography under the conditions of illumination is 4500lx ± 500lx
Detect the purity of glycosides first.
As a result: the appearance of glycosides first is without significant change, and purity is without significant changes.
2, long term test
No. 14004 products are saved under the conditions of room temperature, RH60% ± 10%, are taken respectively at 0,3,6,9,12,18 month
Sample detects glycosides first purity by pharmacopeia chromatography.The results show that the glycosides first content for No. 14004 products just produced is
95.6%, it is saved 12 months under the conditions of room temperature, RH60% ± 10%, purity is without significant changes；It saves 18 months, glycosides first
Purity be 91.7%.
The stability of glycosides first is good, can be reserved for 1.0 years under room temperature, drying regime.
Embodiment
Implementation the present invention relates to glycosides first as treatment herpes simplex virus type 1 and the purposes of 2 types infectious disease, below
Enumerate related embodiment.
1, aqueous solution
1.1 formulas: glycosides first 5g, azone 20g, glycosides first 10g, azone 20g add 10000 mL of deionized water respectively；It is sufficiently molten
Solution, obtains 0.5mg/mL, 1mg/mL glycosides first aqueous solution, filtration sterilization is aseptic subpackaged, every bottle of 5mL.
1.2 application methods:
1.2.1 this product is dipped with sterilized cotton swabs wipe affected part in right amount.Every 2~3h is wiped 1 time, and 5 as a treatment course.Biggish blister
Medication again after rash can be punctured aseptically.
1.2.2 by this product drop on sterile gauze or absorbent cotton, it is applied to herpes infection position, covered plastic film is used
Adhesive tape is fixed.Daily 2~3 times.
1.2.3 as infected in vaginal wall, cervix or ear canal, the sterile gauze or absorbent cotton that have this product will can be dripped
It fills in vagina or ear canal, daily 2~3 times, each 90min.Or this product is dipped with sterilized cotton swabs and wipes affected part, every 4h1 in right amount
Secondary, 7 as a treatment course.
2, tincture
2.1 formulas: glycosides first 5g, azone 20g, glycosides first 10g, azone 20g add 75% medical ethanol 10000mL respectively；Sufficiently
Dissolution, obtains 75% ethanol solution of 0.5mg/mL, 1mg/mL glycosides first, aseptic subpackaged, every bottle of 5mL.
2.2 application methods:
2.2.1 dip this product with sterilized cotton swabs wipes affected part in right amount.Every 2~3h is wiped 1 time, and 5 as a treatment course.Biggish blister
Medication again after rash can be punctured aseptically.
2.2.2 by this product drop on sterile gauze or absorbent cotton, it is applied to herpes infection position, covered plastic film is used
Adhesive tape is fixed.Daily 2~3 times.
3, ointment
3.1. glycosides first beautiful sweetgum resin ointment
3.1.1 it is formulated: glycosides first 1g, beautiful sweetgum resin 25g, vaseline 474g.
3.1.2 preparation method: taking medicine-feeding to be ground into superfine powder, is put into the vaseline of fusing and stirs evenly after mixing.
3.1.3 the major functions: herpe simplex.
3.1.4 usage: affected part is applied when use, 2 times a day, 5 as a treatment course.
3.2 glycosides first beautiful sweetgum resin olibanum myrrh saltcake ointment
3.2.1 it is formulated: glycosides first 1g, beautiful sweetgum resin 25g, frankincense 10g, myrrh 10g, saltcake 10g, vaseline 444g.
3.2.2 preparation method: taking all medicines to be ground into superfine powder, is put into the vaseline of fusing and stirs evenly after mixing.
3.2.3 the major functions: herpe simplex.
3.2.4 usage: by ointment booth on gauze, thick about 5 coin-dividings are affixed on bleb area, with immobilization with adhesive tape.Daily 2
Secondary, 7 as a treatment course.
4, methyl glucoside injection
4.1 formulas: glycosides first 1g adds deionized water 1000mL, obtains 1mg/mL glycosides first aqueous solution, filtration sterilization, and sterile point
Dress, every ampulla 2mL.
4.2 usages: injection (subcutaneous injection), external application (wipe affected part).
5, glycosides first patch (1 patch of Tubeimoside)
5.1 formulas: glycosides first+polyurethane.The patch glycosides first containing Three doses respectively: 5,10,15 milligrams.
5.2 usages: affected part part, a daily patch are affixed on.
Detailed description of the invention
The chemical structure of Fig. 1 Bolbostemma paniculatum glucoside A
Toxicity of Fig. 2 Bolbostemma paniculatum glucoside A to vero cell
Inhibiting effect of Fig. 3 Bolbostemma paniculatum glucoside A to HSV-I/F and HSV-II plaque test
Influence of Fig. 4 Bolbostemma paniculatum glucoside A to HSV-I/F gene duplication
Note: * P < 0.05, significant difference；* P < 0.01, extremely significant difference.
Influence of Fig. 5 Bolbostemma paniculatum glucoside A to HSV-II gene duplication
Note: * P < 0.05, significant difference；* P < 0.01, extremely significant difference.
Claims (1)
1. purposes of the Bolbostemma paniculatum glucoside A as treatment herpes simplex virus I-type and II type infectious disease.
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| CN201711501001.4A CN109953994A (en) | 2017-12-25 | 2017-12-25 | Purposes of the Bolbostemma paniculatum glucoside A as treatment herpes simplex infections disease |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1275378A (en) * | 1999-06-01 | 2000-12-06 | 于廷曦 | Medicinal bait of bolbstemmatoside B for preventing and controlling haman and animal viral disease |
-
2017
- 2017-12-25 CN CN201711501001.4A patent/CN109953994A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1275378A (en) * | 1999-06-01 | 2000-12-06 | 于廷曦 | Medicinal bait of bolbstemmatoside B for preventing and controlling haman and animal viral disease |
Non-Patent Citations (3)
| Title |
|---|
| 张晓辉等: "单纯疱疹病毒性角膜炎土贝母皂甙点眼的浓度筛选与刺激性评价", 《西安交通大学学报(医学版)》 * |
| 张晓辉等: "土贝母皂甙对单疱病毒性角膜炎作用的实验研究", 《眼科新进展》 * |
| 王峰等: "土贝母皂甙对家兔单纯疱疹病毒性角膜炎的作用", 《西安交通大学学报(医学版)》 * |
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