CN102988979A - Artemisia pollen allergen vaccine lozenge and preparation method thereof - Google Patents
Artemisia pollen allergen vaccine lozenge and preparation method thereof Download PDFInfo
- Publication number
- CN102988979A CN102988979A CN2012105415242A CN201210541524A CN102988979A CN 102988979 A CN102988979 A CN 102988979A CN 2012105415242 A CN2012105415242 A CN 2012105415242A CN 201210541524 A CN201210541524 A CN 201210541524A CN 102988979 A CN102988979 A CN 102988979A
- Authority
- CN
- China
- Prior art keywords
- artemisia
- pollen
- pollen allergen
- vaccine
- allergen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides Artemisia pollen allergen vaccine lozenge and a preparation method thereof, wherein the lozenge is prepared by mixing idiotoxins of five kinds of sensitized Artemisia pollens and pharmaceutically acceptable carriers; and the five kinds of sensitized Artemisia plants are Artemisia sieversiana Willd, Artemisia annua L., Artemisia argyi Levl.et Vant, A.ordosica krasch and A.sphaerocep hala krasch. The Artemisia pollen allergen vaccine disclosed by the invention includes idiotoxins of five kinds of main sensitized Artemisia pollens in China; compared with the single Artemisia pollen allergen vaccine, the Artemisia pollen allergen vaccine disclosed by the invention is more effective; furthermore, compared with the sublingual drop, the sublingual lozenge is more steady; and the administration dosage is more accurate.
Description
Technical field
The present invention relates to the allergen vaccine preparation field, specifically, relate to a kind of Artemisia pollen allergen vaccine buccal tablet and preparation method thereof.
Background technology
Human understanding for the allergen disease starts from " Hay Fever ", and scientist noon in 1911 and freeman utilize pollen extracting liquid to treat Hay Fever first, and the allergen disease treatment raises the curtain thus.The development of China's allergen disease treatment is to be introduced from the U.S. by department of otorhinolaryngology director professor Zhang Qingsong of Concord Hospital in 1956 the earliest, and obtained breakthrough, found that Chinese most important autumn type pollen is artemisia (Artemisia) plant pollen, for developing the allergen of this country, realize that the localized road of allergen provides foundation.The eighties was passed the pollen investigation and was shown that what many areas Pollens content summer and autumn was maximum is artimesia by take the lead Chinese gas that tens of the hospitals in the whole nation finish of Ye Shitai, Qiao Bingshan last century.Zhao Jing etc. find in 13～14 years old childrens respiratory tract anaphylactic disease Epidemiological study of Beijing area: the artimesia sensitive subjects is 5.7%.After this, in China each department Artemisia pollen-induced is carried out a large amount of investigation of clinical epidemiology, the result is as follows:
The main allergen that shows Huhehaote City's induced hypersensitivity rhinitis among the Duan Zhijian " Huhehaote City's anaphylactogen distribution situation and clinical analysis " is artimesia.
Zhang Kesheng " area, Huhehaote gas passes the discussion with pollen " result shows that the main pollen in area, Huhehaote is artimesia.
The Meng and Bao Ligao " urban district of Hohhot allergic rhinitis in autumn crowd analysis about allergen detection ": area, Huhehaote main allergen in autumn is sagebruss, and Artemisia apiacea pollen accounts for certain proportion to dirt demodicid mite, DMS person.
Liu Guanghui " research of survey of main allergic pollen in pollinosis in district of Wuhan "; Liu Guanghui, wish that floss flies " Wuhan Area artimesia distribute and on the impact of Allergic Rhinitis airway reactivity ": artimesia is one of Wuhan Urban main pollen in autumn.
Yang Jiong " main pollen investigation in the Wuchang Area air ": artimesia is this area main pollen summer and autumn.
Dong Shangxiong " area, Jingmen, Hubei Province gas passes pollen and allergic asthma dependency ": Jingmen pollen in main autumn is artimesia.
Zhang Hongyu " Beijing's gas passes the investigation of artimesia concentration ": in clinical season, daily mean of concentration is at 100g/m at artimesia in the Beijing area
3
Zhao Yan " application of A Luoge pricking method test in the allergic rhinitis etiological diagnosis ": artimesia is the common anaphylactogens of Jingmen area allergic rhinitis.
Cao Suiping " allergic disease 201 routine sensitinogen analyses ": the main pollen in Yulin Prefecture, Shaanxi Province is artimesia.
Xian Medical Univ's two attached institute Respiratory Medicines " area, Xi'an gas passes the dynamic preliminary survey of pollen ": Xi'an main pollen in area autumn is artimesia.
The Wei Xiu of Second Hospital of Shanxi Medical University blue " Shanxi Province's 1298 example time allergen measurement results are analyzed ": artimesia is Shanxi area childhood asthma allergen principal element.
Datong the 3rd the People's Hospital allergy Ke Li state qin " Datong area 240 routine pollinosis antigen skin test and clinical analysiss ": artimesia tops the list in the Datong area pollen source.
The allergy chamber Zhang Xingyong of hospital general, Lanzhou, the military region " Lanzhou 224 routine allergic rhinitis skin test interpretations of result "; " Lanzhou 87 routine pollinosis analyses "; " the reactive rhinitis investigation report of section of air force mass-sending paraphilia "; The intrathoracic Chen Lin of section of Lanzhou district hospital of army " Lanzhou gas passes the pollen preliminary survey ": the main pollen of Lanzhou is artimesia.
" Luoyang city 360 routine pollinosis patient allergens analyze ": it is artimesia that the main pollen of pollinosis and symptoms of asthma is induced autumn in University Of Science and Technology Of He'nan the 3rd Affiliated Hospital allergy chamber, Foundation of Luoyang.
19 He Hong of smelting hospital precious " Maanshan district 100 routine allergic rhinitis patients' antigen and desensitization observation of curative effect ": the main inhalation (inhalatio) of Maanshan district allergic rhinitis sensitinogen is indoor dust, dust mite, artimesia capsule and polyvalent fungus.
Sun Juan " gas passes the regional investigation of pollen present situation ": all there is Artemisia pollen-induced China eight major regions.
Li Quanwen " gas passes and the pollen investigation in Shandong Province's three city's air ": the main pollen in Jinan City, Shandong Province, Weifang and Yantai is artimesia.
Above artimesia epidemiological investigation shows, Artemisia pollen-induced each area of China that extensively distributes is allergic rhinitis and asthma main pathogenic at the northern area artimesia.
Artemisia (Artemisia) plant is 1-2 or perennial herb, rare shrub, and the normal strong fragrance of tool has root shape neck and nutrition branch more.How upright stem is, and the shape of partly growing thickly has obviously vertical rib; Plant is often by the floss hair; The leaf alternate, most featheriness divisions; Head inflorescence is little, and base portion often has squamella; Spend the opposite sex, the edge is spent female, central authorities' flower both sexes.The pollen characteristic of artemisia has high consistency, and it is spherical in shape or subsphaeroidal to be mainly reflected in pollen, and polar view is three and splits circular tool three hole ditches, and equatorial view is circular or oval.Most chromosome numbers of planting are 2n=18, and the chromosome number of minority kind is 2n=36.It originates in temperate zone, cool temperature zone and the subtropical zone in Asia, Europe and North America.Main irritated artimesia is Artemisia apiacea pollen in North America and Europe, China's artimesia comprises 186 kinds and 44 mutation, spreads all over the country, and northwest, North China, northeast and southwestern provinces and regions are maximum, East China, Central China, south China are less slightly, are born in barren hill, wilderness and roadside more.
China's artimesia distributes the widest, has confirmed clinically to cause the pollen of people's allergy to comprise three kinds: Artemisia apiacea pollen (Artemisia argyi Levl.et Vant), Artemisia sieversiana Willd. pollen (Artemisiasieversiana Wild), Herba Artemisiae annuae pollen (Artemisiaannua L).
The Artemisia injection of the allergic effect Drug Manufacturing Room of Concord Hospital preparation, Herba Artemisiae annuae pollen allergen injection at home hundreds of the hospitals in various places have used decades, effectively suppress the transformation of China's Artemisia pollen-induced rhinitis to allergic asthma, effectively alleviated the symptom of China Artemisia pollen-induced sexually transmitted disease (STD) people asthma.
New Hua Lian, Beijing in 2011 consonance Pharmaceutical is propagated season (July-October) and has been carried out an Epidemiological study at artimesia, to simultaneously pricking method of patient Artemisia apiacea pollen allergen pricking method liquid, Artemisia pricking method liquid, the Herba Artemisiae annuae pollen allergen pricking method liquid of doubtful Artemisia pollen-induced.This time the investigation of clinical epidemiology result shows: in China, there is cross allergy in three kinds of artimesias, although different patient is to three kinds of Artemisia pollen allergen extent of reaction property of there are differences.Also have different types of artimesia in China, existing still the report proves whether other artimesia causes people's allergy.Press the popularity of artimesia distribution, China's artimesia is topmost to be Herba Artemisiae annuae pollen, Artemisia sieversiana Willd. pollen, Artemisia apiacea pollen (seeing " Chinese gas passes pollen and plant color collection of illustrative plates ").Other artimesia is compared this three kinds of artimesias, distribution and energy for growth are relatively little, moreover there is cross immunity originality in Artemisia pollen-induced, Herba Artemisiae annuae pollen, Artemisia sieversiana Willd. pollen, Artemisia apiacea pollen are widely distributed, pollen raw material is easier to collect, so these three kinds of artimesias are suitable material choice of preparation Artemisia pollen allergen vaccine.The Artemisia injection that the allergen Drug Manufacturing Room of BJ Union Hospital produce and Herba Artemisiae annuae pollen allergen injection have proved safe and effective, and the Artemisia apiacea pollen allergen injection in Europe and North America has proved safe and effective.
In recent years, along with the greening propelling in China desert, at the Inner Mongol and Shaanxi one band, Herba artemisiae ordosicae and Folium et Caulis Caryopteridis growth are spread unchecked.Epidemiology in conjunction with China's Artemisia pollen-induced, it is more effective than single Artemisia pollen allergen vaccine that Herba Artemisiae annuae pollen, Artemisia sieversiana Willd. pollen, Artemisia apiacea pollen, Herba artemisiae ordosicae pollen, Folium et Caulis Caryopteridis pollen prepare the Artemisia pollen allergen combined vaccine, the five kinds of more suitable China of Artemisia pollen allergen combined vaccine Artemisia pollen-induced patients' demand, compare the single vaccine of single Artemisia pollen allergen, improved the effectiveness for the treatment of when can guarantee safety.
The method of allergen specific immunity treatment mainly is divided into subcutaneous injection administration immunization therapy and sublingual administration immunization therapy at present.The history in existing 100 years of subcutaneous injection administration immunization therapy, its safety and effectiveness have been obtained proof.At the beginning of the nineties in last century, allergen Sublingual drop vaccine is born, and 1998, WHO announced allergen Sublingual drop vaccine safety and effective.The beginning of this century, Sublingual drop allergen vaccine is widely used in Europe, South America, Australia, Southeast Asia and Middle East.Sublingual drop allergen vaccine is compared subcutaneous injection administration allergen vaccine, and is safer, patient's medication of can going home voluntarily, and patient compliance is good, is easy to accept this therapeutic modality.In allergic rhinitis and the asthma patient, child patient accounts for significant proportion, for the children population patient, and Sublingual immunity drop allergen vaccine, patient compliance is better, acceptant this therapeutic modality.
Yet a shortcoming of Sublingual drop vaccine is, the dosage that at every turn splashes into is difficult to control, and the dosage inaccuracy differs greatly, and preparation allergen vaccine buccal tablet is more stable with respect to the Sublingual drop, can realize the homogenization administration.
Summary of the invention
The purpose of this invention is to provide a kind of Artemisia pollen allergen vaccine buccal tablet and preparation method thereof.
In order to realize the object of the invention, the invention provides a kind of Artemisia pollen allergen vaccine, described vaccine comprises the allergen of five kinds of sensitization artimesias, and five kinds of sensitization sagebrusses are: Artemisia sieversiana Willd. (Artemisia sieversiana Willd), Herba Artemisiae annuae (Artemisia annua L.), Radix Artemisia ordosicae (Artemisia argyi Levl.et Vant), Herba artemisiae ordosicae (A.ordosica krasch) and Folium et Caulis Caryopteridis (A.sphaerocep hala krasch).
Aforesaid Artemisia pollen allergen vaccine, the allergenic activity ratio of five kinds of sensitization artimesias is preferably 1:1:1:1:1.
The preparation method of above-mentioned Artemisia pollen allergen vaccine may further comprise the steps:
1) gather by machinery (vacuum cleaner) or water culture is collected five kinds of artimesias, cross respectively 200～300 mesh sieves and remove impurity such as dust in the pollen, and pollen is carried out purity detecting, and morphology and molecular biology identification.
2) pollen after the remove impurity is used the defat of acetone soaking and stirring continuously, with the solids natural drying after the defat.
3) pollen behind the extracting degreasing adds in sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution of 50～500mM with the rate of charge of 1:10～50(g:mL) respectively, and 4 ℃ are stirred 4～24h.
4) with gained solution in the step 3) successively through 8 μ m, 0.8 μ m, 0.45 μ m and 0.22 μ m membrane filtration.
5) with the ultrafilter membrane weighing apparatus filter of gained filtrate in the step 4) with molecular cut off 5kD, until that the filter liquor supernatant is the electrical conductivity of colourless or filter liquor is consistent with the used buffer of ultrafiltration, namely get each allergen vaccine stock solution.
6) compete the activity that immunodepression is measured each vaccinogen liquid with RSAT, then be mixed in proportion each vaccinogen liquid, namely get the Artemisia pollen allergen vaccine.
The present invention also provides a kind of Artemisia pollen allergen vaccine buccal tablet, is comprised of above-mentioned Artemisia pollen allergen vaccine and pharmaceutically acceptable carrier.
Aforesaid Artemisia pollen allergen vaccine buccal tablet, the percentage by weight of each component is:
Wherein, Artemisia pollen allergen vaccine: mannitol: crosslinked Carboxymethyl cellulose sodium: anhydrous silica gel: microcrystalline Cellulose: magnesium stearate: the weight ratio of Lactose hydrate is preferably 1:5:1:0.02:0.1:0.05:0.01.
The present invention also provides the preparation method of above-mentioned Artemisia pollen allergen vaccine buccal tablet, may further comprise the steps:
1) gather by machinery (vacuum cleaner) or water culture is collected five kinds of artimesias, cross respectively 200～300 mesh sieves and remove impurity such as dust in the pollen, and pollen is carried out purity detecting, and morphology and molecular biology identification.
2) pollen after the remove impurity is used the defat of acetone soaking and stirring continuously, with the solids natural drying after the defat.
3) pollen behind the extracting degreasing adds in sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution of 50～500mM with the rate of charge of 1:10～50(g:mL) respectively, and 4 ℃ are stirred 4～24h.
4) with gained solution in the step 3) successively through 8 μ m, 0.8 μ m, 0.45 μ m and 0.22 μ m membrane filtration.
5) with the ultrafilter membrane weighing apparatus filter of gained filtrate in the step 4) with molecular cut off 5kD, until that the filter liquor supernatant is the electrical conductivity of colourless or filter liquor is consistent with the used buffer of ultrafiltration, namely get each allergen vaccine stock solution.
6) compete the activity that immunodepression is measured each vaccinogen liquid with RSAT, then be mixed in proportion each vaccinogen liquid, namely get the Artemisia pollen allergen vaccine.
7) add lyophilizing after the mannitol in the Artemisia pollen allergen vaccine.
8) powder behind the Artemisia pollen allergen vaccine freeze-drying and crosslinked Carboxymethyl cellulose sodium, anhydrous silica gel, microcrystalline Cellulose, magnesium stearate and Lactose hydrate are mixed in proportion rear tabletting, vacuum packaging and get final product.
Artemisia pollen allergen vaccine sublingual lozenge provided by the invention comprises three concentration specifications: 50000AU/ sheet, 5000AU/ sheet, 500AU/ sheet.In use comprise two stages of dosage escalation and maintenance dose.
The present invention further provides the application that described Artemisia pollen allergen vaccine and Artemisia pollen allergen vaccine sublingual lozenge are used for the anaphylactic disease of specific active immunotherapy Artemisia pollen-induced mediation.Described anaphylactic disease comprises: allergic dermatitis, chronic urticaria, allergic rhinitis and allergic asthma etc.
In the present invention, described Artemisia is to separate the Artemisia that obtains from Artemisia sieversiana Willd. pollen, described Herba Artemisiae annuae pollen allergen is to separate the Herba Artemisiae annuae pollen allergen that obtains from Herba Artemisiae annuae pollen, described Artemisia apiacea pollen allergen is to separate the Artemisia apiacea pollen allergen that obtains from Artemisia apiacea pollen, described Herba artemisiae ordosicae pollen allergen is to separate the Artemisia apiacea pollen allergen that obtains from Herba artemisiae ordosicae pollen, and described Folium et Caulis Caryopteridis Artemisia pollen allergen is to separate the Folium et Caulis Caryopteridis pollen allergen that obtains from Folium et Caulis Caryopteridis pollen.
Five kinds of selected Artemisia pollen allergen of Artemisia pollen allergen vaccine sublingual lozenge of the present invention's preparation: Artemisia, Herba Artemisiae annuae pollen allergen, Artemisia apiacea pollen allergen, Herba artemisiae ordosicae pollen allergen and Folium et Caulis Caryopteridis pollen allergen are five kinds of Artemisia pollen allergen of the main sensitization of China, and its effectiveness is higher than single Artemisia pollen allergen.The vaccine of five kinds of Artemisia pollen allergen associatings has been included the main allergen of China's sensitization artimesia.The relative allergen vaccine of allergen vaccine sublingual lozenge Sublingual drop is more convenient for transporting, stable storage, and each dosage is consistent, has improved the effectiveness that uses.
The specific embodiment
Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.If do not specialize, the conventional means that used technological means is well known to those skilled in the art among the embodiment, the raw materials used commercial goods that is.
The preparation (50000AU/ sheet) of embodiment 1 Artemisia pollen allergen vaccine sublingual lozenge
Prepare respectively Herba Artemisiae annuae pollen allergen, Artemisia, Artemisia apiacea pollen allergen, Herba artemisiae ordosicae pollen allergen and Folium et Caulis Caryopteridis pollen allergen stock solution: gather or five kinds of artimesias of water culture collection by machinery (vacuum cleaner), cross respectively 200～300 mesh sieves and remove impurity such as dust in the pollen, and pollen carried out purity detecting, and morphology and molecular biology identification.Pollen after the remove impurity is used the defat of acetone soaking and stirring continuously, with the solids natural drying after the defat.Take by weighing five kinds of each 100g of artimesia pollen after the defat with rate of charge 1:25(g:mL) add in 50mM sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution, 4 ℃ are stirred 24h.With gained solution with 8 μ m membrane filtrations, again with filtrate successively through 0.8 μ m, 0.45 μ m, 0.22 μ m membrane filtration.Gained filtrate is with the ultrafilter membrane weighing apparatus filter of molecular cut off 5kD, until that the filter liquor supernatant is the electric conductance of colourless or filter liquor is consistent with buffer with ultrafiltration, namely gets each allergen vaccine stock solution.Measure the activity of each vaccinogen liquid with RSAT competition immunodepression.Mix the rear 4% mannitol lyophilizing of adding by specific activity 1:1:1:1:1, the powder after the lyophilizing (being Artemisia pollen allergen vaccine freeze-drying powder), every gram contains Artemisia pollen allergen 100,0000AU.
By following prescription supplementary material is mixed:
After above supplementary material mixes, fully stir, behind the mix homogeneously, 10000 of the Artemisia pollen allergen sublingual lozenge vaccines of employing direct compression legal system, every 0.1g (containing Artemisia pollen allergen 50000AU).
The preparation (5000AU/ sheet) of embodiment 2 Artemisia pollen allergen vaccine sublingual lozenges
Prepare Herba Artemisiae annuae pollen allergen, Artemisia, Artemisia apiacea pollen allergen, Herba artemisiae ordosicae pollen allergen and Folium et Caulis Caryopteridis pollen allergen stock solution according to the method for describing among the embodiment 1, mix the rear 4% mannitol lyophilizing of adding by specific activity 1:1:1:1:1, powder after the lyophilizing (being Artemisia pollen allergen vaccine freeze-drying powder), every gram contains Artemisia pollen allergen 100000AU.
By following prescription supplementary material is mixed:
After above supplementary material mixes, fully stir, behind the mix homogeneously, 10000 of the Artemisia pollen allergen sublingual lozenge vaccines of employing direct compression legal system, every 0.1g (containing Artemisia pollen allergen 5000AU).
The preparation (500AU/ sheet) of embodiment 3 Artemisia pollen allergen vaccine sublingual lozenges
Prepare Herba Artemisiae annuae pollen allergen, Artemisia, Artemisia apiacea pollen allergen, Herba artemisiae ordosicae pollen allergen and Folium et Caulis Caryopteridis pollen allergen stock solution according to the method for describing among the embodiment 1, mix the rear 4% mannitol lyophilizing of adding by specific activity 1:1:1:1:1, powder after the lyophilizing (being Artemisia pollen allergen vaccine freeze-drying powder), every gram contains Artemisia pollen allergen 10000AU.
By following prescription supplementary material is mixed:
After above supplementary material mixes, fully stir, behind the mix homogeneously, 10000 of the Artemisia pollen allergen sublingual lozenge vaccines of employing direct compression legal system, every 0.1g (containing Artemisia pollen allergen 500AU).
Synergism in 4 five kinds of Artemisia pollen allergen pharmacodynamicss of embodiment
1.1 animal sensitization
With reference to the people such as Hsiue TR, Int Arch Allergy Immuol1997; 112 (3): the method for describing among the 295-302 is adsorbed onto 1%Al (OH) with 20mg Artemisia, Herba Artemisiae annuae pollen allergen, Artemisia apiacea pollen allergen, Herba artemisiae ordosicae pollen allergen, Folium et Caulis Caryopteridis pollen allergen mixed solution
3On the gel, give Cavia porcellus subcutaneous injection 0.5ml, in addition five kinds of Artemisia pollen allergen of 20mg are dissolved in 0.1%Al (OH)
3On the gel, give same guinea pig intraperitoneal injection 0.5ml.Every interval was repeated sensitization 1 time, totally 3 times in 10 days.
1.2 administering mode
It is 8 groups that test group is divided into: normal group, matched group, five kinds of artimesia mixing allergen administration groups, Artemisia administration group, Herba Artemisiae annuae pollen allergen administration group, Artemisia apiacea pollen allergen administration group, Herba artemisiae ordosicae pollen allergen administration group, Folium et Caulis Caryopteridis pollen allergen administration group.Normal group is in not administration of duration of test, and matched group is the allergen solvent.The specification (embodiment 3) of five kinds of artimesia mixing allergen tyrosine Adjuvanted vaccines: 1000AU/ml(4 number), 100AU/ml(3 number), 10AU/ml(2 number), 1AU/ml(1 number).
Tyrosine adjuvant Artemisia vaccine, tyrosine adjuvant Herba Artemisiae annuae pollen allergen vaccine, tyrosine adjuvant Artemisia apiacea pollen allergen vaccine, tyrosine adjuvant Herba artemisiae ordosicae pollen allergen vaccine, tyrosine Folium et Caulis Caryopteridis pollen allergen vaccine specification: 1000AU/ml(4 number), 100AU/ml(3 number), 10AU/ml(2 number), 1AU/ml(1 number).Every group is cleaned 10 of level sensitized guinea pigs.
Consumption: every group of Cavia porcellus subcutaneous injection, inject once weekly, administration time is 6 months altogether.No. 1 medicine: in the 1st～4 week, inject respectively weekly 0.1ml, 0.2ml, 0.4ml, 0.8ml.
No. 2 medicines: in 5～8 weeks, inject respectively weekly 0.1ml, 0.2ml, 0.4ml, 0.8ml.No. 3 medicines: in the 9th～12 week, inject respectively weekly 0.1ml, 0.2ml, 0.4ml, 0.8ml.No. 4 medicines: in the 13rd～16 week, inject respectively weekly 0.1ml, 0.2ml, 0.4ml, 0.8ml.Since the 17th week, inject weekly medicine 1ml No. 4, to off-test.
1.3 pharmacodynamic evaluation
After administration finished, every group of Cavia porcellus was respectively at diverse location subcutaneous injection 0.5ml100AU/ml Artemisia aqueous solution, Herba Artemisiae annuae pollen allergen aqueous solution, Artemisia apiacea pollen allergen aqueous solution, Herba artemisiae ordosicae pollen allergen aqueous solution, Folium et Caulis Caryopteridis pollen allergen aqueous solution.Measure the welt of every Cavia porcellus different injection sites generation and the diameter (meansigma methods of major diameter and minor axis diameter) of erythema, calculate the arithmetical average of every kind of pollen allergen welt of every group of Cavia porcellus and erythema diameter.The results are shown in Table 1.
Five kinds of Artemisia pollen allergen vaccines of table 1 are worked in coordination with pharmacodynamic evaluation
Annotate: 1 group of administration: be five kinds of Artemisia pollen allergen mixed vaccines of tyrosine adjuvant; 2 groups of administrations: be tyrosine adjuvant Artemisia epidemic disease Artemisia sieversiana Willd. pollen vaccine; 3 groups of administrations: be tyrosine adjuvant Herba Artemisiae annuae pollen allergen vaccine; 4 groups of administrations: tyrosine adjuvant Artemisia apiacea pollen allergen vaccine; 5 groups of administrations: tyrosine adjuvant Herba artemisiae ordosicae pollen allergen vaccine; 6 groups of administrations: tyrosine adjuvant Folium et Caulis Caryopteridis pollen allergen vaccine.
As known from Table 1: single tyrosine adjuvant Artemisia pollen allergen vaccine, can effectively reduce corresponding Artemisia pollen-induced degree, also can reduce simultaneously other four kinds of Artemisia pollen-induced degree, but the relative five kinds of artimesia mixing allergen vaccines of single tyrosine Artemisia pollen allergen vaccine, its significant degree to the treatment of other four kinds of Artemisia pollen-induced is mixed tyrosine vaccine (p＜0.01) not as five kinds of Artemisia pollen allergen.Result by this experiment has proved that there is intercrossing in five kinds of Artemisia pollen allergen, but the also property of there are differences, the effectiveness of five kinds of Artemisia pollen allergen combined vaccines is higher than single Artemisia pollen allergen vaccine.
The detection of embodiment 5 Artemisia pollen allergen vaccine sublingual lozenge activity
Artemisia pollen allergen Sublingual drop vaccine activity detects the method for using UniCAP to adopt the experiment of RAST competitive inhibition.
50000AU/ sheet, 5000AU/ sheet, 500AU/ sheet are respectively got 10, use physiological saline solution, the finished product of 3 specifications and internal reference product are 10 with normal saline dilution respectively
-1, 10
-2, 10
-3, 10
-4, 10
-5Then totally 5 concentration mix with 100 μ l equivalent serum, hatch 60min for 37 ℃, then to the upper machine testing of Artemisia apiacea pollen allergen immunoCAP, calculate suppression ratio, take the logarithm value of dilution factor 1/10 as abscissa, percent inhibition is vertical coordinate, the dilution factor when calculating 50% suppression ratio.Dilution factor by 50% suppression ratio is compared with the internal reference product, calculates activity (AU).The results are shown in Table 2.
Table 2 three batch sample activity values
The investigation of embodiment 6 Artemisia pollen allergen vaccine sublingual lozenge stability
This experiment is passed through three batches of Artemisia pollen allergen vaccine sublingual lozenges 2-8 ℃ of placement, March, June, JIUYUE, December, active variation in 18 months, the stability of investigation Artemisia pollen allergen vaccine sublingual lozenge (table 3-table 5).
Table 3201006001 batch stability experiment data
Table 4201007001 batch stability experiment data
Table 5201008001 batch stability experiment data
Above experimental result shows, Artemisia pollen allergen vaccine sublingual lozenge three batch samples were placed 18 months at 2-8 ℃, actively significantly do not change, this condition place 18 months basicly stable.
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (7)
1. Artemisia pollen allergen vaccine, it is characterized in that, described vaccine comprises the allergen of five kinds of sensitization artimesias, and five kinds of sensitization sagebrusses are: Artemisia sieversiana Willd. (Artemisiasieversiana Willd), Herba Artemisiae annuae (Artemisia annua L.), Radix Artemisia ordosicae (Artemisia argyiLevl.et Vant), Herba artemisiae ordosicae (A.ordosica krasch) and Folium et Caulis Caryopteridis (A.sphaerocephala krasch).
2. vaccine according to claim 1 is characterized in that, the allergenic activity of five kinds of sensitization artimesias is than being 1:1:1:1:1.
3. the preparation method of claim 1 or 2 described vaccines is characterized in that, may further comprise the steps:
1) gather five kinds of artimesias, mistake 200～300 mesh sieves are removed the impurity in the pollen respectively;
2) pollen after the remove impurity soaks with acetone, stirs defat, drying;
3) pollen after the defat is added in sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution of 50～500mM with the rate of charge of 1:10～50(g:mL) respectively, 4 ℃ are stirred 4～24h;
4) with gained solution in the step 3) successively through 8 μ m, 0.8 μ m, 0.45 μ m and 0.22 μ m membrane filtration;
5) with the ultrafilter membrane weighing apparatus filter of gained filtrate in the step 4) with molecular cut off 5kD, until that the filter liquor supernatant is the electrical conductivity of colourless or filter liquor is consistent with the used buffer of ultrafiltration, namely get each allergen vaccine stock solution;
6) compete the activity that immunodepression is measured each vaccinogen liquid with RSAT, then be mixed in proportion each vaccinogen liquid, and get final product.
4. an Artemisia pollen allergen vaccine buccal tablet is characterized in that, is comprised of claim 1 or 2 described Artemisia pollen allergen vaccines and pharmaceutically acceptable carrier.
5. buccal tablet according to claim 4 is characterized in that, the percentage by weight of each component is:
6. buccal tablet according to claim 5, it is characterized in that Artemisia pollen allergen vaccine: mannitol: crosslinked Carboxymethyl cellulose sodium: anhydrous silica gel: microcrystalline Cellulose: magnesium stearate: the weight ratio of Lactose hydrate is 1:5:1:0.02:0.1:0.05:0.01.
7. the preparation method of claim 5 or 6 described Artemisia pollen allergen vaccine buccal tablets is characterized in that, may further comprise the steps:
1) gather five kinds of artimesias, mistake 200～300 mesh sieves are removed the impurity in the pollen respectively;
2) pollen after the remove impurity soaks with acetone, stirs defat, drying;
3) pollen after the defat is added in sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution of 50～500mM with the rate of charge of 1:10～50(g:mL) respectively, 4 ℃ are stirred 4～24h;
4) with gained solution in the step 3) successively through 8 μ m, 0.8 μ m, 0.45 μ m and 0.22 μ m membrane filtration;
5) with the ultrafilter membrane weighing apparatus filter of gained filtrate in the step 4) with molecular cut off 5kD, until that the filter liquor supernatant is the electrical conductivity of colourless or filter liquor is consistent with the used buffer of ultrafiltration, namely get each allergen vaccine stock solution;
6) compete the activity that immunodepression is measured each vaccinogen liquid with RSAT, then be mixed in proportion each vaccinogen liquid, namely get the Artemisia pollen allergen vaccine;
7) add lyophilizing after the mannitol in the Artemisia pollen allergen vaccine;
8) powder behind the Artemisia pollen allergen vaccine freeze-drying and crosslinked Carboxymethyl cellulose sodium, anhydrous silica gel, microcrystalline Cellulose, magnesium stearate and Lactose hydrate being mixed in proportion rear tabletting makes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210541524.2A CN102988979B (en) | 2012-12-13 | 2012-12-13 | Artemisia pollen allergen vaccine lozenge and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210541524.2A CN102988979B (en) | 2012-12-13 | 2012-12-13 | Artemisia pollen allergen vaccine lozenge and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102988979A true CN102988979A (en) | 2013-03-27 |
| CN102988979B CN102988979B (en) | 2014-05-07 |
Family
ID=47918413
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210541524.2A Active CN102988979B (en) | 2012-12-13 | 2012-12-13 | Artemisia pollen allergen vaccine lozenge and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102988979B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106967696A (en) * | 2017-02-22 | 2017-07-21 | 杭州艾乐吉生物科技有限公司 | A kind of anaphylactogen of Artemisia Plant Pollen and its application |
| CN111175512A (en) * | 2018-11-09 | 2020-05-19 | 北京新华联协和药业有限责任公司 | Preparation and test of artemisia pollen specific IgE antibody standard serum |
| WO2022028069A1 (en) * | 2020-08-03 | 2022-02-10 | 格林生物医药科技(天津)有限公司 | Allergen preparation |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108265064B (en) * | 2016-12-31 | 2020-06-09 | 江苏众红生物工程创药研究院有限公司 | Recombinant artemisia annua 3-class allergen protein and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004067660A (en) * | 2002-08-02 | 2004-03-04 | Coletica | TYPE 2 OR TYPE 3 HUMAN beta-DEFENSIN-STIMULATING ACTIVE COMPONENT, AND COSMETIC AND MEDICINAL COMPOSITION CONTAINING THE SAME |
| CN1785288A (en) * | 2005-11-11 | 2006-06-14 | 鹰潭医药有限责任公司 | Oral disintegration tablet for treating cough ant phlegm and its preparation method |
| CN101496901A (en) * | 2008-02-03 | 2009-08-05 | 浙江我武生物科技有限公司 | Application of acidic buffer in stable pollen allergen activity |
| CN101791399A (en) * | 2009-02-04 | 2010-08-04 | 阿尔克-阿贝洛有限公司 | Water-containing liquid allergic reaction bacterin preparation for mouth mucosa drug administration |
| CN101905022A (en) * | 2009-06-08 | 2010-12-08 | 浙江我武生物科技有限公司 | Stable medicinal composition containing artemisia pollen allergen and preparation method thereof |
| CN102552900A (en) * | 2011-12-21 | 2012-07-11 | 北京新华联协和药业有限责任公司 | Artemisia pollen allergen vaccine, and preparation method and application thereof |
-
2012
- 2012-12-13 CN CN201210541524.2A patent/CN102988979B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004067660A (en) * | 2002-08-02 | 2004-03-04 | Coletica | TYPE 2 OR TYPE 3 HUMAN beta-DEFENSIN-STIMULATING ACTIVE COMPONENT, AND COSMETIC AND MEDICINAL COMPOSITION CONTAINING THE SAME |
| CN1785288A (en) * | 2005-11-11 | 2006-06-14 | 鹰潭医药有限责任公司 | Oral disintegration tablet for treating cough ant phlegm and its preparation method |
| CN101496901A (en) * | 2008-02-03 | 2009-08-05 | 浙江我武生物科技有限公司 | Application of acidic buffer in stable pollen allergen activity |
| CN101791399A (en) * | 2009-02-04 | 2010-08-04 | 阿尔克-阿贝洛有限公司 | Water-containing liquid allergic reaction bacterin preparation for mouth mucosa drug administration |
| CN101905022A (en) * | 2009-06-08 | 2010-12-08 | 浙江我武生物科技有限公司 | Stable medicinal composition containing artemisia pollen allergen and preparation method thereof |
| CN102552900A (en) * | 2011-12-21 | 2012-07-11 | 北京新华联协和药业有限责任公司 | Artemisia pollen allergen vaccine, and preparation method and application thereof |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106967696A (en) * | 2017-02-22 | 2017-07-21 | 杭州艾乐吉生物科技有限公司 | A kind of anaphylactogen of Artemisia Plant Pollen and its application |
| CN106967696B (en) * | 2017-02-22 | 2020-01-14 | 杭州艾乐吉生物科技有限公司 | Allergen of artemisia pollen and application thereof |
| CN111175512A (en) * | 2018-11-09 | 2020-05-19 | 北京新华联协和药业有限责任公司 | Preparation and test of artemisia pollen specific IgE antibody standard serum |
| WO2022028069A1 (en) * | 2020-08-03 | 2022-02-10 | 格林生物医药科技(天津)有限公司 | Allergen preparation |
| CN114053402A (en) * | 2020-08-03 | 2022-02-18 | 格林生物医药科技(天津)有限公司 | Allergen preparation |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102988979B (en) | 2014-05-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101905022B (en) | Stable medicinal composition containing artemisia pollen allergen and preparation method thereof | |
| CN102552900A (en) | Artemisia pollen allergen vaccine, and preparation method and application thereof | |
| CN102988979B (en) | Artemisia pollen allergen vaccine lozenge and preparation method thereof | |
| CN101195647B (en) | Hydroxyl carthamus tinctorius yellow colour A, preparation method and application thereof | |
| Zhi-Gang et al. | A study on pollen allergens in China | |
| CN101310751B (en) | Detection method of medicine composition for replenishing qi and blood | |
| US20110059124A1 (en) | The quality control method and application of a kind of ganoderma lucidum spore oil fat emulsion | |
| CN100457139C (en) | Method for preparing a Shuanhuanglian injection and the component detecting method | |
| CN102512672B (en) | Artemisia pollen allergen vaccine and preparation method thereof | |
| CN101128736A (en) | Method for monitoring patient or subject compliance with medical prescriptions, and formulation for use in the method | |
| CN101869656B (en) | Chinese medicinal preparation for treating cough with asthma and preparation method thereof | |
| CN100400075C (en) | Qi-invigorating, blood-nourishing medicinal composition and its preparing method | |
| Frostad et al. | Clinical effects of hyposensitization using a purified allergen preparation from Timothy pollen as compared to crude aqueous extracts from Timothy pollen and a four‐grass pollen mixture respectively | |
| CN102512673B (en) | Injectable artemisia pollen allergen vaccine and preparation method thereof | |
| CN102258742B (en) | Chinese medicinal medicine composition for treating depression and preparation method thereof | |
| CN1281268C (en) | Chinese medicinal composition for supplementing Qi, nourishing heart and calming mind and its preparation and identification | |
| CN104922564A (en) | Preparation method of traditional Chinese medicine pills for treating tuberculosis | |
| CN109939227A (en) | A kind of ragweed pollen allergen extract, its immersion liquid and preparation method thereof | |
| CN102327341A (en) | Oral solution for abating fever of cold in children and preparation method thereof | |
| CN115400194A (en) | Traditional Chinese medicine composition for treating childhood epidemic and application thereof | |
| CN1299734C (en) | Composition of traditional Chinese medicine for large intestine hygropyretic disease and its preparation method | |
| CN101670088B (en) | Traditional Chinese medicine composite for treating acute/chronic bronchitis | |
| CN1714840A (en) | Chinese medicine composition for stopping itching and its preparing method | |
| CN102579869A (en) | Preparation method of traditional Tibetan medicine composition for treating liver diseases | |
| CN101904894A (en) | Application of lamiophlomis rotate total glycosides to preparing medicines |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |