CN1275378A - Medicinal bait of bolbstemmatoside B for preventing and controlling haman and animal viral disease - Google Patents
Medicinal bait of bolbstemmatoside B for preventing and controlling haman and animal viral disease Download PDFInfo
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Abstract
The present invention relates to a bolbostemma glucoside B, bolbostemma saponin containing bolbostemma glucoside B and its application for preparing medicine or medicinal bait for preventing and curing human and animal various viral diseases. The bolbostemma glucoside B is a kind of chemical component separated from bolbostemma, and the bolbostemma saponin is the saponin component in the bolbostemma containing no bolbostemma glucoside C. The bolbostemma glucoside B and bolbostemma saponin can inhibit the replication of human and animal virus, so that it can be usecd for preventing and curing human and animal various viral diseases, and can obtain good therapeutic effect.
Description
The present invention relates to bolbstemmatoside B and contain the Rhizoma Bolbostematis saponin preparation prevention of bolbstemmatoside B and the medicine or the pharmaceutical chemistry of the various viral diseases of treatment humans and animals.
As everyone knows, virus is a kind of serious harm human health, causes that multiple animal suffers from various illness even dead microorganism virulence factor.Pandemic prawn disease viral disease and bird flu that human health is threatened very big acquired immune deficiency syndrome (AIDS), various influenza, various viral hepatitis, dust Po Shi viral infection and causes tremendous economic to lose to the mankind causes by virus institute in the whole world at present.Some common virus, as herpes simplex virus (HSV) though the infection that causes usually is asymptomatic, but it not only can cause local patholoic change, as the genitals of outbreak repeatedly and damage, keratitis, the conjunctivitis of actinal surface portion, also can cause serious systemic disease, as meningitis and neonate whole body disseminated infections etc.Other common virus, then relevant with the generation of cervical cancer and nasopharyngeal carcinoma respectively as papillomavirus (HPV) with Epstein-Barr (EB) virus.It should be noted that some zoosis toxicity disease, can also infect to the people.For various viral diseases, the mankind still do not have the effectively preventing means at present.
The natural derivative fritillary bulb methyl glucoside injection of having described bolbstemmatoside B in Chinese patent description (CN 1094637A) has duplicated the obvious suppression effect to human AIDS poison I type and variant thereof, and can suppress the caused cytopathy of human AIDS poison I type.Above-mentioned patent application at this as a reference.
Bolbstemmatoside B is a kind of monomer that separation and Extraction is come out from the Chinese medicine Rhizoma Bolbostematis, and molecular weight is 1334, and chemical structural formula as shown in Figure 1.The difference of bolbstemmatoside B and Lobatoside H is: on 16 carbon of Lobatoside H aglycon is hydrogen, and is hydroxyl on 16 carbon of bolbstemmatoside B aglycon.Bolbstemmatoside B is the same with Lobatoside H, good water solubility, and stability is strong, and only yield is than Lobatoside H low (bolbstemmatoside B is 0.5%, and Lobatoside H is 1.8%).Rhizoma Bolbostematis saponin involved in the present invention means the saponin component in the Rhizoma Bolbostematis that does not contain Rhizoma Bolbostematis glycosides third, and its water solublity and stability are then suitable with Lobatoside H, bolbstemmatoside B.Prepare the simple for process of Rhizoma Bolbostematis saponin, yield is also high, and about 2.2%.
The extracting method of bolbstemmatoside B and Rhizoma Bolbostematis saponin is referring to document (Wang Yongqing, Yu Lijian etc.: Shaanxi new medicine, 10:55,1981; Kasai, R et al., Phytochemistry, 27:1439-1446,1988)
The present invention relates to bolbstemmatoside B or Rhizoma Bolbostematis saponin as prevention and the medicine of the various viral diseases of treatment humans and animals or the compositions of pharmaceutical chemistry, this medicine or pharmaceutical chemistry compositions comprise bolbstemmatoside B or Rhizoma Bolbostematis saponin and pharmaceutically acceptable or edible carrier.
The present invention relates to as defined medicine of claim 1 or pharmaceutical chemistry preparation of compositions method, this method comprises that bolbstemmatoside B or Rhizoma Bolbostematis saponin mix with pharmaceutically acceptable or edible carrier respectively or dissolve.
The present invention relates to bolbstemmatoside B or Rhizoma Bolbostematis saponin and carry out application among the human or animal of this prevention or treatment at needs as the medicine of prevention and treatment humans and animals various viral diseases or pharmaceutical chemistry.
Bolbstemmatoside B or Rhizoma Bolbostematis saponin can mix or dissolve with any pharmaceutically acceptable or edible carrier makes pharmaceutical composition, and these carriers comprise in skin, mucosa, gastrointestinal and pharmaceutically suitable carrier of parenteral or edible carrier.This pharmaceutical composition uses with the form of conventional pharmaceutical formulation, as the tablet of solid form, granule, powder, capsule, cachet, suppository etc., or the ointment of semi-solid form, ointment etc., or injection of liquid form, suspending agent, syrup, Emulsion, liniment etc.Use pharmaceutically useful excipient and additive in this pharmaceutical composition, these pharmaceutically useful excipient and additive comprise nontoxic compatible filler, binding agent, disintegrating agent, buffer agent, antiseptic, antioxidant, lubricant, correctives, thickening agent, coloring agent, emulsifying agent, stabilizing agent, for example lactose, citric acid, stearic acid, magnesium stearate Gypsum Fibrosum powder, sucrose, corn starch, Pulvis Talci, gelatin, agar, pectin, Oleum Arachidis hypogaeae semen, cocoa butter, ethylene glycol, glucose, procaine hydrochloride, lidocaine hydrochloride, ascorbic acid etc.This pharmaceutical composition can be by the common process preparation of various preparations.This pharmaceutical chemistry compositions is used with the form of conventional bait, uses pharmaceutically acceptable or edible excipient and additive in the bait composition.
Bolbstemmatoside B that reaches of the present invention or Rhizoma Bolbostematis saponin are as the medicine of the various viral diseases of prevention humans and animals, can be used as food additive, cosmetics additive, vagina and join in food, cosmetics, contraceptive or the contraceptive cover, also can join in the living environment of bait or aquatic animal with contraceptive or contraceptive cover additive, additive of bait.
Active ingredient bolbstemmatoside B of the present invention or Rhizoma Bolbostematis saponin be as the medicine of the various human and animal virus's property diseases of control, can be used for all kinds of illness, acquired immune deficiency syndrome (AIDS) and prawn ' s virus disease that various viral hepatitis, herpes simplex virus cause, bird flu etc.
The content of bolbstemmatoside B or Rhizoma Bolbostematis saponin is the about 0.001-100mg of every kg body weight every day in the made various combination of oral medication, preferably about 0.01-50mg, more excellent about 0.05-30mg, best about 0.1-10mg.Divide every day and take for 2-4 time, 5-30 days is a course of treatment, and general medication 3-6 course of treatment, each, the interval was 5-30 days course of treatment.Made medicinal composition for injections can supply intramuscular injection or intravenous drip, and wherein the content of bolbstemmatoside B or Rhizoma Bolbostematis saponin is every kg body weight 0.001-50mg every day, preferably about 0.01-25mg/kg, more excellent about 0.05-15mg, best about 0.1-3mg.Divide 1-3 use every day, 5-30 days is a course of treatment, and general medication 3-6 course of treatment, each, the interval was 5-30 days course of treatment.Yet accurate dose, route of administration, application method etc. should determine by the doctor, depends primarily on age, body weight, the state of an illness, reaction of patient etc.As for animal-use drug, then should do suitably to adjust to the tolerance and the response situation of medicine according to animal.
Enumerate our experiment content and result thereof below, can be used for preparing prevention and treat the medicine of various human and animal virus's property diseases and the evidence of pharmaceutical chemistry as bolbstemmatoside B or Rhizoma Bolbostematis saponin.One. pharmacodynamics test
1. bolbstemmatoside B or Rhizoma Bolbostematis saponin are to the inhibitory action of herpes simplex virus I-type, II type (HSV-I, HSV-II)
1.1. cell: Vero-E
6Cell strain (African green monkey kidney cell strain).
1.2. virus: HSV-I and HSV-II, titre is respectively PFU (plaque forming unit 5 * 10
6/ ml, 4 * 10
8/ ml, working concentration is MOI=1 in the experiment.
1.3. bolbstemmatoside B or Rhizoma Bolbostematis saponin are in external neutralization to herpes simplex virus I-type and II type.
1.4. virus is inoculated after bolbstemmatoside B or the effect of Rhizoma Bolbostematis saponin: the medicinal liquid of variable concentrations is mixed with equal-volume herpes simplex virus liquid, act on and add in the cell monolayer hole every hole 0.1ml, the parallel work of each concentration 3 holes after 1.5 hours respectively.Day by day observation of cell pathological changes situation.Establish virus control group, variable concentrations medicinal liquid matched group and normal control group simultaneously.
1.5. add bolbstemmatoside B or Rhizoma Bolbostematis saponin after the cell infection virus: in growing up to the hole of cell monolayer, add herpes simplex virus, 0.1ml/ hole, the parallel work of each concentration 3 holes.Discard viral liquid after in 37 ℃, the incubator of 5% carbon dioxide, saturated humidity, hatching 1.5 hours.Wash bolbstemmatoside B or the Rhizoma Bolbostematis saponin that adds variable concentrations after 3 times respectively, observation of cell pathological changes situation day by day with culture fluid.Establish virus control group, variable concentrations medicinal liquid matched group and normal control group simultaneously.
1.6. result
1.6.1 bolbstemmatoside B or Rhizoma Bolbostematis saponin have direct deactivation to HSV-I type and HSV-II type, the results are shown in Table 1,2.
Table 1 bolbstemmatoside B is to the external neutralization of HSV-I
*
Observing time
72h 96h
The cell contrast------
Virus control ++ ++ ++ ++ ++ ++ ++ ++ ++ ++ ++ ++
Bolbstemmatoside B
(μg/ml)
0.5 - - -
1.0 - - -
2.0 small part cavity
4.0 part cavity
8.0 it is dead
Virus+soil
Bulbus Fritillariae Uninbracteatae glycosides second
(μg/ml)
0.5 ++++ ++++ ++++ ++++ ++++ ++++
1.0 ++++ ++++ ++++ ++++ ++++ ++++
2.0 ++ ++ ++ +++ +++ +++
4.0 + + + ++ ++ ++
8.0 - - -
*-cell pathological changes ± individual cells do not occur and pathological changes, shrinkage occur
Pathological changes appears in+25% following cell ++ and 25-50% cell circle contracts, pathological changes
+++50-75% cell circle contracts, pathological changes ++ ++ the external neutralization of pathological changes table 2 bolbstemmatoside B to HSV-II appears in 75% above cell
Observing time
72h 120h
The cell contrast------
Virus control ++ ++ ++ ++ ++ ++ ++ ++ ++ ++ ++ ++
Bolbstemmatoside B
(μg/ml)
0.5 - - -
1.0 - - -
2.0 small part cavity
4.0 part cavity
8.0 it is dead inflexibly
Virus+soil
Bulbus Fritillariae Uninbracteatae glycosides second
(μg/ml)
0.5 ++++ ++++ ++++ ++++ ++++ ++++
1.0 ++ +++ ++ ++++ ++++ ++++
2.0 + + + +++ ++ ++
4.0 ± - - + ± ±
8.0 - - -
*-cell pathological changes ± individual cells do not occur and pathological changes, shrinkage occur
Pathological changes appears in+25% following cell ++ and 25-50% cell circle contracts, pathological changes ++ and+50-75% cell circle contracts, pathological changes ++ ++ pathological changes appears in 75% above cell
1.6.2 bolbstemmatoside B or Rhizoma Bolbostematis saponin have protective effect to HSV-I type and HSV-II type cytopathogenic effect, the results are shown in Table 3,4.Table 3 bolbstemmatoside B is to the cytopathogenic protective effect of HSV-I
*
The 72h observed result
The cell contrast---
Virus control ++ ++ ++ ++ ++ ++
The Rhizoma Bolbostematis glycosides
The second contrast
(μg/ml)
0.5 - - -
1.0 - - -
2.0 small part cavity
4.0 part cavity
8.0 it is dead
Virus+soil
Bulbus Fritillariae Uninbracteatae glycosides second
(μg/ml)
0.25 ++++ ++++ ++++
0.5 ++++ ++++ ++++
1.0 ++++ ++++ ++++
2.0 +++ +++ +++
4.0 ++ ++ ++
8.0 - - -
*-cell pathological changes ± individual cells do not occur and pathological changes, shrinkage occur
Pathological changes appears in+25% following cell ++ and 25-50% cell circle contracts, pathological changes
+++50-75% cell circle contracts, pathological changes ++ ++ 75% above cell pathological changes table 4 bolbstemmatoside B occurs to the cytopathogenic protective effect * of HSV-II
Observing time
72h 120h
The cell contrast------
Virus control ++ ++ ++ ++ ++ ++ ++ ++ ++ ++ ++ ++
Bolbstemmatoside B
(μg/ml)
0.5 - - - - - -
1.0 - - - - - -
2.0 small part cavity
4.0 part cavity
8.0 it is dead
Virus+soil
Bulbus Fritillariae Uninbracteatae glycosides second
(μg/ml)
0.5 ++++ ++++ ++++ ++++ ++++ ++++
1.0 ++++ ++++ ++++ ++++ ++++ ++++
2.0 ++ +++ ++ +++ +++ +++
4.0 + + + ++ ++ ++
8.0 - - -
*-cell pathological changes ± individual cells do not occur and pathological changes, shrinkage occur
Pathological changes appears in+25% following cell ++ and 25-50% cell circle contracts, pathological changes
+++50-75% cell circle contracts, pathological changes ++ ++ pathological changes appears in 75% above cell
1.7 presentation of results
With the herpesvirus infection cell behind the 2-4 μ g/ml medicinal liquid effect certain hour, the cytopathy degree is starkly lower than matched group.After infection the 5th day, virus control group cell almost all came off, and became the garden by swelling and contracted and HSV-II experimental group pathological changes shows as cell, and refractivity weakens, and the part cell breakage does not but come off or seldom comes off.The above results shows that bolbstemmatoside B has direct deactivation to HSV-I type and HSV-II type.In addition, the Rhizoma Bolbostematis saponin slightly is inferior to bolbstemmatoside B (data does not show) to the direct deactivation of HSV-I type and HSV-II type.
In the experiment of medication, bolbstemmatoside B concentration has retarding action to the herpesvirus cytopathogenic effect to elder generation's infective virus when 2-4 μ g/ml then.Experiment also shows, when same concentration, medication person's cytopathy time of occurrence is prior to neutralization group (medicinal liquid and viral interaction in vitro postoperative infection groups of cells) behind elder generation's infective virus, and the cytopathy degree shows that also than neutralization group height the bolbstemmatoside B medicinal liquid is bigger than its inhibitory action to the direct deactivation of virus.The effect that the Rhizoma Bolbostematis saponin suppresses HSV-I type and HSV-II type slightly is inferior to bolbstemmatoside B (data does not show).
2. bolbstemmatoside B or Rhizoma Bolbostematis saponin are to the inhibitory action of HIV (human immunodeficiency virus) (HIV-I)
2.1. cell: T lymphoblastoid strain (MT-2 cell), to contain the RPMI-1640 culture medium culturing of 10% hyclone.
2.2. virus: the chorista HTLV-III of HIV-I
B, HTLV-III
RFAnd HTLV-III
MNWith H9 and MT-2 cell culture.Virus is by centrifugal (150 * g), 8 μ m aperture membrane filtration culture supernatant acquisitions.The titre of virus storage liquid is 1 * 10
5-10 * 10
5(every milliliter of MT-2 cell 50% tissue infection dosage, TCID
50).
2.3. the virus neutralization is measured: bolbstemmatoside B or Rhizoma Bolbostematis saponin are measured with following two kinds of methods the neutralization activity that HIV-I infects: suppress core protein P
24The generation of (the nucleocapsid antigen of HIV-I) and protection cell exempt from HIV-I pathological changes caused by virus (colorimetry).Bolbstemmatoside B and Rhizoma Bolbostematis saponin do not contain phenol red RPMI-1640 culture fluid and carry out system dilution to contain 10% hyclone.Diluent adds 96 orifice plates and is that 0.0015 HIV-I mixes with the infection multiplicity (MOI) of equivalent dilution.Cultivate after 1 hour for 37 ℃, add the MT-2 cell (1 * 10 of 25 μ l polybrene (1,5-dimethyl-1,5-phenodiazine 11 methylene gather Methobromide) processing
6/ ml).After hatching under 37 ℃ of conditions, from each hole, collect 100 μ l culture supernatant, replenish the fresh medium of equivalent simultaneously in each hole.Supernatant is used to measure P
24Each hole added XTT tetrazole indicator (1mg/ml, 50 μ l/ holes) in the 6th day.Under the 150nm condition, measure intracellular first after 4 hours.HIV duplicates or cytopathy inhibition percentage rate is calculated as follows: (C-B)/C * 100, B and C represent P when adding or not adding bolbstemmatoside B or Rhizoma Bolbostematis saponin respectively
24Amount or cytotoxic percentage rate.
2.4 result
2.4.1 bolbstemmatoside B suppresses HTLV-III
BReplication-competent virus core protein P
24Measure with the ELISA method.Bolbstemmatoside B suppresses P
24Generation be the dose-effect dependence.Bolbstemmatoside B suppresses HTLV-III
BThe EC that duplicates
50Be 21.6 μ g/ml.
2.4.2. bolbstemmatoside B suppresses the cytopathy bolbstemmatoside B of HIV mediation and obviously suppresses HTLV-III
BThe cytopathy of mediation.Bolbstemmatoside B suppresses HTLV-III
BThe cytopathic 50% valid density (EC of mediation
50) be 20.4 μ g/ml.
2.4.3. the inhibition HTLV-III that bolbstemmatoside B duplicates other HIV-I separated strains
BIt is one of separated strain of the most normal use in the experiment.Bolbstemmatoside B also suppresses HTLV-III effectively
MNAnd HTLV-III
RFDuplicate its EC
50Be respectively 21.8 μ g/ml and 18.1 μ g/ml.The inhibitory action that bolbstemmatoside B duplicates the various separated strains of HIV-1 slightly is better than Lobatoside H (referring to patent CN 1094637A), and the activity of Rhizoma Bolbostematis saponin slightly is inferior to bolbstemmatoside B but be better than Lobatoside H (data does not show).
2.4.4. the cell toxicant of bolbstemmatoside B mediation when the concentration of bolbstemmatoside B when 80 μ g/ml are diluted to 40 μ g/ml, its cytotoxicity reduces rapidly.50% cell toxicant concentration (CC of bolbstemmatoside B
50) be 62.8 μ g/ml.The Rhizoma Bolbostematis saponin is to HTLV-III
BThe inhibition of duplicating, to the inhibition that other HIV-I separated strains duplicate, the cytopathic inhibition effect that HIV is mediated all slightly is inferior to bolbstemmatoside B, and its cytotoxicity then slightly is better than bolbstemmatoside B (data does not show).Lobatoside H slightly is inferior to the Rhizoma Bolbostematis saponin to the inhibition effect that HIV-I duplicates, and cytotoxicity is slightly strong than it.
3. bolbstemmatoside B or Rhizoma Bolbostematis saponin are to Epstein-Barr virus and the collaborative inhibition effect of bringing out nasopharyngeal carcinoma of TPA (12-0-myristoyl Fo Bo-13-acetas)
(Nasopharyngeal Carcinoma NPC) has substantial connection to Epstein-Barr virus (EBV) with nasopharyngeal carcinoma.Change the PSG-CR2-Hyg carrier in the immortal human epithelial cell (293 cell) acquisition CR2-293 cell.Indirect immunofluorescence is measured and is found, cell transformed is expressed the Epstein-Barr virus receptor.Behind ebv infection CR2-293 cell, 1/4 cell can be expressed the antigen of Epstein-Barr virus.After acting on these cells with TPA, the cell number of expressing virus antigen increases.From the DNA of ebv infection cell, amplify epstein barr virus dna W segment with the PCR method.Under the TPA continuous action, the morphological characteristic of ebv infection cell changes.Subcutaneous the ebv infection cell inoculation nude mice, inject TPA weekly, inducing cell forms tumor in nude mouse.Through tissue pathology checking, tumor is low differentiation cell carcinoma (Li Baomin etc., viral journal, 14:133-138,1998).In the surrounding injection TPA of ebv infection cell inoculation position, inject bolbstemmatoside B or Rhizoma Bolbostematis saponin weekly, and observe tumor formation situation every day.The result shows, behind the ebv infection CR2-293 cell, adds 5ng/mlTPA effect to the during 4 weeks, and cellular morphology changes.Some cells are in heaps, the multilamellar growth, and the contact inhibition growth is lost, and the cell feeler tails off, shortens.Behind the ebv infection CR2-293 cell, it is subcutaneous to be inoculated in nude mice, weekly cell inoculation position injection 50ng/mlTPA or in injection TPA at same area injection bolbstemmatoside B or Rhizoma Bolbostematis saponin.After 3 weeks, the cell inoculation position lump occurs and grows up gradually.6-7 is during week, and lump is quite obvious.Experiment shows, the about half of organizing except that the subcutaneous TPA of adding of ebv infection CR2-293 cell inoculation of nude mice occurs the tumor, other each groups, promptly inoculate the ebv infection cell but do not inject TPA, or injection TPA time injection bolbstemmatoside B or Rhizoma Bolbostematis saponin, or inoculation tumor do not occur without the nude mice of injection TPA group behind 293 cells of ebv infection.This shows that bolbstemmatoside B or Rhizoma Bolbostematis saponin have the inhibition effect of highly significant to Epstein-Barr virus and the collaborative induced tumor of TPA.
4. bolbstemmatoside B or Rhizoma Bolbostematis saponin are to the therapeutic effect of rabbit herpetic keratitis
4.1. bolbstemmatoside B or Rhizoma Bolbostematis saponin eye drop: prepare under aseptic condition with normal saline, making the content of bolbstemmatoside B wherein or Rhizoma Bolbostematis saponin is 1mg/ml.
4.2. 0.1% acycloguanosine eye liquid (production of Qianjiang, Hubei pharmaceutical factory).
4.3. virus: herpes simplex virus I-type SM
44Strain (HSV-SM
44), titre is 10
6Median tissue culture infective dose (TCID
50)/ml draws from Beijing Biological Product Inst..
4.4. animal: 24 of healthy new zealand rabbits, body weight 2-3kg, male and female half and half.
4.5. infection model and experiment grouping: after cornea is done 4mm# word impression, splash into virus stock solution used 30 μ l, massage 30s closes one's eyes.Be divided into 4 groups at random, i.e. bolbstemmatoside B treatment group, Rhizoma Bolbostematis saponin treatment group, acycloguanosine eye liquid positive controls and normal saline blank group are numbered 1,2,3,4 groups respectively, and each organizes the lagophthalmos number average is 12.
4.6. eye drip treatment: capable eyes administration in 24 hours behind the virus inoculation, every each 2 of eye (about 50 μ l), every day 5 times, continuous use 14 days.
4.7. observation of curative effect: every day the row fluorescence staining, slit lamp observation record keratopathy is divided into the 0-4 level with the corneal epithelium lesion degree by the Trousdale method.
4.8. result: 24-28h behind the inoculation HSV-1, point-like, short dendroid pathological changes all appear in all test rabbit corneals, and each organizes the mutual comparing difference of keratopathy degree does not have significance (P＞0.05).The expansion that keeps of matched group inoculation back the 3rd day, these typical dendroid focuses of cornea; Formed large stretch of intensive dendroid and map shape ulcer in 6-8 days, the substrate edema is soaked into, and the pathological changes area accounts for the 75%-100% of the cornea gross area; Pathological changes was expanded to deep layer in 9-14 days, and epithelium ulcer area is the trend of dwindling.Bolbstemmatoside B or Rhizoma Bolbostematis saponin treatment group after inoculation the 3rd day, visible point-like of cornea and corynebacterium pathological changes, scope is limitation.When the state of an illness was heavier to 4-6 days, the pathological changes area accounted for the 20%-40% of cornea area.The substrate edema, soak into also gentlyer, highly significant (P＜0.01) is arranged with matched group keratopathy degree comparing difference.Fully recover to the 12nd day most of rabbit cornea.Acycloguanosine eye liquid treatment group, after inoculation 4-6 days, the pathological changes area accounted for the 25%-45% of cornea area.Most of rabbit cornea recovery from illness to the 14th day.The ascending order of therapeutic effect is: acycloguanosine eye liquid＜Rhizoma Bolbostematis saponin＜bolbstemmatoside B.
5. bolbstemmatoside B or Rhizoma Bolbostematis saponin are to the therapeutic effect of dermopathic herpesvirus disease
Dermopathic herpesvirus disease system is by due to the human simple herpesvirus (Herpes virus hominis).This viral nucleic acid is DNA.Herpes simplex virus can be divided into I, II type (being called for short HSV-I and HSV-II).HSV-I is relevant with most of non-genital infection, and HSV-II infects and mostly occurs at genital area, finds that recently HSV-II is relevant with the morbidity of cervical cancer.Dermopathic herpesvirus disease can be divided into herpes simplex, chickenpox, herpes zoster, Kaposi varicelliform eruption, cytomegalovirus infection, infectious monocytosis, B virosis etc.Bolbstemmatoside B or Rhizoma Bolbostematis saponin all have certain therapeutic effect to above-mentioned dermopathic herpesvirus disease.Now with recurrence type herpes labialis (herpes facialis) (herpes labiais; Herpesfacialis) therapeutic effect of 30 examples is an example.Herpes labialis is a modal type in the recurrent herpes simplex, that outbreak initial stage part often has earlier is scorching hot, pruritus and flushing, then occur in groups intensive or the several groups of big phlysises of syringe needle, want little and than a small bundle of straw, etc. for silkworms to spin cocoons on collection than the vesicle of essential, rotten to the corn sepage after breaking, dry gradually incrustation, in overall process 1-2 week, more temporary pigmentation can be left in the part, back.Damage is good to be sent out in mucocutaneous intersection, near bicker, lip edge and nostril, also betides face, lip person.This organizes 30 examples, male 18 examples, and women 12 examples, age distribution is at 18-65 between year.Scorching hot, pruritus and flare phase 8 examples; Vesicle phases 12 example; Vesicle is broken, rotten to the corn phases 10 example.The bolbstemmatoside B of usage: 5mg/ml or Rhizoma Bolbostematis saponin aqueous solution are smeared the part, and 1 hour 1 time, 30 routine patients all fully recover in day at 1-3, more the local non-pigment calmness in back.90% patient was not recurred in 1 year.Therapeutic effect Rhizoma Bolbostematis saponin to primary disease slightly is inferior to bolbstemmatoside B.
6. bolbstemmatoside B or Rhizoma Bolbostematis saponin are to papova viruses treatment of diseases effect papova viruses (papovaviruses), belong to DNA viruses, be human papillomavirus (papillomavirus), polyoma virus (palyoma virus) and vacuolating virus (Simiare vacnolatingvirus, SV
40).Human papillomavirus (HPV) not only causes wart after infecting, and some can cause malignant tumor, as skin carcinoma, carcinoma of tongue and cervical cancer etc.Bolbstemmatoside B or Rhizoma Bolbostematis saponin are better to the verruca plana (claim not only flat wart) and the therapeutic effect of genital wart (but also claiming condyloma acuminatum).The bolbstemmatoside B of usage: 5mg/ml or Rhizoma Bolbostematis saponin aqueous solution are smeared the part, every day 8-12 time.Non-stimulated to the part, do not stay cicatrix after healing.To the therapeutic effect of primary disease, the Rhizoma Bolbostematis saponin slightly is inferior to bolbstemmatoside B.
7. bolbstemmatoside B or Rhizoma Bolbostematis saponin are women's a kind of commonly encountered diseases, frequently-occurring disease to the therapeutic effect chronic cervicitis of chronic cervicitis, relevant with papillomavirus and HIV-II, show as cervical erosion, plumpness, polyp etc. clinically, some cases brings out cervical cancer.Bolbstemmatoside B or Rhizoma Bolbostematis saponin are made suppository, insert the cervix uteri part, not only chronic cervicitis are had excellent curative, and can also prevent the generation of cervical cancer, and the partial cancer in situ of cervix uteri is taken a turn for the worse.To the therapeutic effect of chronic cervicitis, the Rhizoma Bolbostematis saponin slightly is inferior to bolbstemmatoside B, but is better than Lobatoside H (referring to patent CN 1110133 A).
8. since the eighties, world's shrimp farming develops rapidly as the purposes of preparation control prawn ' s virus disease medicament and pharmaceutical chemistry for bolbstemmatoside B or Rhizoma Bolbostematis saponin, and output is from several ten thousand tons of more than 70 ten thousand tons of being increased to the nineties at the beginning of the eighties.When aquaculture develops rapidly, also broken out global prawn disease, caused serious harm to aquaculture.Only 1993-1994 China prawn disease outbreak of epidemic just makes aquaculture first mate's degree underproduction of China, with a toll of billions of units.It has been established that, and prawn disease fulminant in the world's is popular is in recent years mainly caused by viral infection.Have now found that prawn virus disease has kind more than 20, what cause disease of prawn in recent years mainly is baculovirus (a kind of DNA viruses).Free length different incubation period behind the prawn infective virus, be the inapparent infection state.Behind the prawn infective virus in case morbidity, promptly progress rapidly, symptom such as just occur in 1-2 day not ingesting is difficult to treatment.Prawn ' s virus is very sensitive to bolbstemmatoside B or Rhizoma Bolbostematis saponin.Bolbstemmatoside B or Rhizoma Bolbostematis saponin are low to prawn and mammiferous toxicity.Bolbstemmatoside B and Rhizoma Bolbostematis saponin are all soluble in water, and room temperature is long following stabilization time, and is good through gastrointestinal absorption.Make bait feeding with bolbstemmatoside B or Rhizoma Bolbostematis saponin, can prevent not infect or the prawn morbidity of inapparent infection state.If shrimp falls ill, mix use with oxytetracycline, prawn disease there is certain therapeutic effect.To the prevention effect of prawn ' s virus disease, the Rhizoma Bolbostematis saponin slightly is inferior to bolbstemmatoside B (data does not show).Two. toxicological test
1. acute toxicity test selects for use the ICR mice to test with muscle and intraperitoneal injection method.Result of the test: the LD of intramuscular injection bolbstemmatoside B
50Be 45.66mg/Kg, the LD of Rhizoma Bolbostematis saponin
50Be 42.36mg/Kg.The LD of the bolbstemmatoside B of intraperitoneal administration
50Be 20.9mg/Kg, the LD of Rhizoma Bolbostematis saponin
50Be 19.6mg/Kg.
2. the toxic action to normal cultured cell adopts colorimetry to test the toxic action of bolbstemmatoside B to the T-Lymphoblastoid strain (MT-2 cell) of In vitro culture, result of the test (seeing Table 5) shows, when the concentration of bolbstemmatoside B is reduced to 40 μ g/ml by 80 μ g/ml, cytotoxicity significantly descends, when concentration was lower than 40 μ g/ml, toxicity was very low.The cytotoxic CD of bolbstemmatoside B
50Be 68 μ g/ml, the cytotoxic CD of Lobatoside H
50Be 59 μ g/ml.The cytotoxic CD of Rhizoma Bolbostematis saponin
50Be 64 μ g/ml.Table 5 bolbstemmatoside B is to the toxic action of MT-2 cell
Concentration (μ g/ml) cytotoxic percentage rate (%) bolbstemmatoside B 10 1.5
20 2.0
40 10.0
80 84.6 Lobatoside H 10 2.2
20 6.5
40 13.0
80 96.4
Fig. 1: the chemical structural formula of Lobatoside H, glycosides second and glycosides third
Three. embodiment
Following formula examples has illustrated dosage form of the present invention.
Embodiment 1: tablet
The NO amounts of components
Mg/ sheet mg/ sheet
1 bolbstemmatoside B (or Rhizoma Bolbostematis saponin) 50 100
2 lactose USP 122 123
3 corn starchs, food stage is 30 40
10% pure water slurry
4 corn starchs, food stage 95 130
5 magnesium stearate 37
Add up to 300 400
Manufacture method
The 1st and the 2nd component of mixing is 15 minutes in suitable mixer, the 3rd component and mixture granulation.If desired, grind moist granule, and make it dry by a primary dcreening operation.If desired, dried granules is sieved, and mixed 10-15 minute with the 4th.Adding the 5th mixed 1-3 minute.In suitable tablet machine, mixture is pressed into suitable size and weight.
Embodiment 2: capsule
The NO amounts of components
Mg/ grain mg/ grain
1 bolbstemmatoside B (or Rhizoma Bolbostematis saponin) 50 100
2 lactose USP 106 120
3 corn starchs, food stage 40 73
4 magnesium stearate NF 47
Add up to 200 300
Manufacture method
The 1st, 2 and the 3rd were mixed 10-15 minute in suitable mixer, add the 4th and mixed 1-3 minute.On suitable encapsulation machine with in the capsule that it is suitable that this mixture is packed into.
Embodiment 3: injection
Bolbstemmatoside B (or Rhizoma Bolbostematis saponin) 10g or 50g
Glucose 500g
Water for injection adds to 10000ml
Manufacture method
Get bolbstemmatoside B (or Rhizoma Bolbostematis saponin), glucose is dissolved in an amount of 60 ℃ water for injection, adds 0.05% injection-use activated carbon by amount of preparation and stirs, and leaves standstill 15 minutes, filter.Adding the injection water, to make final volume be 10000ml, stirs evenly.Sampling and measuring pH value and content, qualified after-filtration, 115 ℃ of pressure sterilizings of embedding 30 minutes, promptly.
Embodiment 4: suppository is example with 10000 of production invention products, and used raw material and proportioning thereof are: bolbstemmatoside B (or Rhizoma Bolbostematis saponin) 300g or 600g Tween 80 800g cacao bean ester (or semi-synthetic fatty acid glycerine) add to the 10000g manufacture method
Mix: the proportioning by present embodiment takes by weighing bolbstemmatoside B (or Rhizoma Bolbostematis saponin), Tween 80 and cacao bean ester (or semi-synthetic fatty acid glycerine), cacao bean ester (or semi-synthetic fatty acid glycerine) is put into stainless-steel pan heating in water bath to major part dissolve, stop heating.Add Tween 80 and bolbstemmatoside B (or Rhizoma Bolbostematis saponin) then respectively, be stirred to mix homogeneously, just obtain hybrid medicine of the present invention with the rustless steel blender.
The preparation of lubricant:, make lubricant with green soap and each abundant mixing and stirring of 80% medical alcohol a and 5 times of glycerol.
Molding: on the bolt mould, coat the lubricant of having prepared.Hybrid medicine of the present invention is poured in the bolt mould to overflowing die orifice, allowed its natural cooling.After treating to solidify fully, cut with cutter and to overflow part.Opening mold takes out suppository from mould then.
Aseptic packaging: quality examination is carried out in the prepared agent of fastening, and every weight should be 1g, contains bolbstemmatoside B (or Rhizoma Bolbostematis saponin) 30mg or 60mg.Pack cobalt 60 sterilizations after the passed examination.
Embodiment 5: eye drop bolbstemmatoside B (or Rhizoma Bolbostematis saponin) 1g ethyl hydroxybenzoate 0.3g sodium chloride injection adds to the 1000ml manufacture method
Get ethyl hydroxybenzoate and be dissolved in an amount of thermal chlorination sodium injection, put coldly, filter.Adding the chlorination sodium injection, to make volume be 1000ml, 100 ℃ of flowing steam sterilizations 30 minutes, and it is standby to make aseptic ethyl hydroxybenzoate sodium chloride solution.Get bolbstemmatoside B (or Rhizoma Bolbostematis saponin) by aseptic manipulation, be dissolved in an amount of aseptic ethyl hydroxybenzoate sodium chloride solution, make near 950ml.Take out a little and measure PH (transferring PH to 5.0-7.0 with the 0.1mol/L caustic lye of soda in case of necessity).Add the ethyl hydroxybenzoate sodium chloride solution again and make into 1000ml, mixing.Filter in case of necessity, promptly aseptic subpackaged.This product contains bolbstemmatoside B (or Rhizoma Bolbostematis saponin) and should be 0.09-0.11% (g/ml).
Embodiment 6: Eye ointments bolbstemmatoside B (or Rhizoma Bolbostematis saponin) 1g eye pasting substrate adds to the 1000g manufacture method
Eye pasting substrate prescription: lanoline 100g, sterile liquid paraffin 100g, vaseline 800g.Get lanoline and vaseline and put heat fused in the suitable container with lid.Add sterilized liquid paraffin, mixing, filtered while hot in case of necessity, 150 ℃ of dry heat sterilizations 1 hour are put cold standby.Get bolbstemmatoside B (or Rhizoma Bolbostematis saponin) by aseptic manipulation, put in the sterilization mortar, add a small amount of sterilized liquid paraffin and grind, make into fine and smooth pasty state, gradation adds eye pasting substrate again.The limit edged grinds well, and makes into 1000g, and is aseptic subpackaged.This product contains bolbstemmatoside B (or Rhizoma Bolbostematis saponin) and should be 0.09-0.11% (g/g).
Embodiment 7: ointment bolbstemmatoside B (or Rhizoma Bolbostematis saponin) 10g lanoline 90g vaseline 900g manufacture method
Get bolbstemmatoside B (or Rhizoma Bolbostematis saponin) by aseptic manipulation, the lanoline, the vaseline that add an amount of fusing grind, and gradation adds remaining lanoline, vaseline again, makes into 1000g, grinds well promptly.
Embodiment 8: nasal drop bolbstemmatoside B (or Rhizoma Bolbostematis saponin) 5g ephedrine hydrochloride 10g glycerol 300ml distilled water adds to the 1000ml manufacture method
Get bolbstemmatoside B (or Rhizoma Bolbostematis saponin) by aseptic manipulation and be dissolved in the hot distilled water, add ephedrine hydrochloride and make moltenly, filter glycerol adding.Stir evenly, add water to the 1000ml mixing, packing, promptly.
Embodiment 9: syrup bolbstemmatoside B (or Rhizoma Bolbostematis saponin) 2g distilled water 20ml simple syrup adds to the 1000ml manufacture method
Get bolbstemmatoside B (or Rhizoma Bolbostematis saponin) by aseptic manipulation, add in the 20ml distilled water and dissolve.Add simple syrup to 1000ml, stir evenly, packing, promptly.
Embodiment 10: solution bolbstemmatoside B (or Rhizoma Bolbostematis saponin) 50g sterile distilled water adds to the 10000ml manufacture method
Get bolbstemmatoside B (or Rhizoma Bolbostematis saponin) by aseptic manipulation, add the dissolving of 10000ml sterile distilled water, packing, promptly.During use, calculate the dose that adds of per kilogram feedstuff by the body weight of fish and shrimp.If fish and shrimp are fallen ill, then should mix use with oxytetracycline.
Claims (3)
1. prevent and treat the pharmaceutical composition of the various viral diseases of humans and animals, this pharmaceutical composition comprises bolbstemmatoside B or Rhizoma Bolbostematis saponin and pharmaceutically useful carrier.
2. as the defined preparation of drug combination method of claim 1, this method comprises that bolbstemmatoside B or Rhizoma Bolbostematis saponin mix with pharmaceutically useful carrier respectively or dissolve.
3. bolbstemmatoside B or Rhizoma Bolbostematis saponin are used for preventing and treating the purposes of the various viral disease medicines of humans and animals in manufacturing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB99107839XA CN1147302C (en) | 1999-06-01 | 1999-06-01 | Medicinal bait of bolbstemmatoside B for preventing and controlling haman and animal viral disease |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB99107839XA CN1147302C (en) | 1999-06-01 | 1999-06-01 | Medicinal bait of bolbstemmatoside B for preventing and controlling haman and animal viral disease |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1275378A true CN1275378A (en) | 2000-12-06 |
| CN1147302C CN1147302C (en) | 2004-04-28 |
Family
ID=5272977
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB99107839XA Expired - Fee Related CN1147302C (en) | 1999-06-01 | 1999-06-01 | Medicinal bait of bolbstemmatoside B for preventing and controlling haman and animal viral disease |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1147302C (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109260247A (en) * | 2018-10-23 | 2019-01-25 | 渭南新秦药物研究有限责任公司 | A kind of skin antiseptic and preparation method thereof acting on herpesviral |
| CN109953994A (en) * | 2017-12-25 | 2019-07-02 | 于立坚 | Purposes of the Bolbostemma paniculatum glucoside A as treatment herpes simplex infections disease |
-
1999
- 1999-06-01 CN CNB99107839XA patent/CN1147302C/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109953994A (en) * | 2017-12-25 | 2019-07-02 | 于立坚 | Purposes of the Bolbostemma paniculatum glucoside A as treatment herpes simplex infections disease |
| CN109260247A (en) * | 2018-10-23 | 2019-01-25 | 渭南新秦药物研究有限责任公司 | A kind of skin antiseptic and preparation method thereof acting on herpesviral |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1147302C (en) | 2004-04-28 |
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