CN101926994B - Turtle shell immunoadjuvant and influenza vaccine containing the same - Google Patents
Turtle shell immunoadjuvant and influenza vaccine containing the same Download PDFInfo
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Abstract
The invention relates to a turtle shell immunoadjuvant and an influenza vaccine containing the immunoadjuvant; the turtle shell immunoadjuvant comprises the following ingredients in percentage by weight: 0.001-100% of turtle shell extractive and 0-99.999% of carriers. The composition of the influenza vaccine containing the turtle shell immunoadjuvant is as follows: the mass ratio of the turtle shell immunoadjuvant to the influenza vaccine hemagglutinin is 50-180050. The turtle shell immunoadjuvant in the invention has a good immunoadjuvant effect and has an effect of effectively preventing diseases in human and animal vaccine. The turtle shell extractive is used as the immunoadjuvant of the influenza vaccine; compared with the traditional aluminum hydroxide immunoadjuvant, the turtle shell immunoadjuvant has high using safety and small byeffect, can well improve the original nature of the influenza vaccine; in the process of preparing the influenza vaccine, the method is simple, economic and environment friendly and convenient; the method easily controls the mass, greatly reduces the dosage of the influenza vaccine antigen and satisfies the immunity demand when occurring the epidemic influenza.
Description
Technical field
The present invention relates to a kind of Carapax Trionycis immunological adjuvant, also relate to the influenza vaccines that contain the Carapax Trionycis immunological adjuvant, belong to biological technical field,
Technical background
Carapax Trionycis is the carapace of Trionychidae animal Trionyx sinensis Wiegmann (Trionyx sinensis Wiegmann), main product in Hubei, ground such as Hunan, Anhui, it is very wide to distribute; By the Hainan Island in northeast to Guangdong, and ground such as Sichuan, Yunnan, Shaanxi, Gansu all has, and all can catch the whole year; With autumn, two seasons of winter be many, kill in the rearmounted boiling water to scald when sclerderm can peel off to carapace and take out, remove residual meat; Dry, or use with quench in vinegar behind the stir-baking with sand.The Carapax Trionycis beginning is stated from Shennong's Herbal, and the successive dynasties book on Chinese herbal medicine is all on the books.Property is salty, is slightly cold, and returns liver, kidney channel." control the labor evil spirit, remove bone heat " carried in another name for Sichuan Province this " property of medicine opinion "." book on Chinese herbal medicine converge with speech " carries " Carapax Trionycis removes the hot malaria of the deficiency of YIN, and the medicine of separating the consumptive fever hectic fever due to YIN-deficiency also ", can control a little less than the yang deficiency stomach, and the food drink does not disappear, and vomiting and nausea is had loose bowels." Japan hanako materia medica " carry " with pyrexial malaria, abdominal mass due to blood stasis, dyspepsia, cold, mass beside the umbilicus, cold abdominal mass and broken crux serve as speech also ".Be that " Chinese pharmacopoeia is recorded kind to version in 2005, has nourishing YIN for suppressing the hyperactive YANG, and hard masses softening and resolving is brought down a fever and removed the effect of steaming.Be mainly used in fever due to yin deficiency, consumptive fever hectic fever due to YIN-deficiency, stirring-up of pathogenic wind in the interior resulting from deficiency, amenorrhea ， WEIJIA, diseases such as chronic malaria malaria with splenomegaly.
Year people have carried out a lot of researchs to its chemical constituent and pharmacotoxicological effect aspect surplus in the of nearly ten, experiment showed, that Carapax Trionycis extract has can obviously improve mice hypoxia-bearing capability and anti-refrigeration, prolong the mice swimming time, have antifatigue effect; Can significantly improve mice plaque forming cells's haematolysis ability, promote hemolytic antibody to generate; And can strengthen the mice delayed hypersensitivity.Can increase muscle contraction intensity and the effect of drawing the stricture of vagina area, prolonging lasting contraction time.Have the hypertrophy that suppresses connective tissue, the effect that improves plasma protein, mass dissipating and swelling eliminating, can improve the antineoplastic immunocompetence, can reduce experimental hyperthyroidism animal blood slurry CAMP content; Can improve the lymphoblastic transformation rate, prolong the antibody life period, raise immunity; Can protect adrenal cortex function; Can promote hemopoietic function, improve content of hemoglobin; Can suppress connective tissue proliferation, so the lump that can dissipate; Prevent the cell mutation effect; Also has certain sedation.
The plurality of active ingredients that these pharmacotoxicological effects and Carapax Trionycis are contained, closely related like compositions such as animal glue, ossein, keratin, Trionyx sinensis Wiegmann polysaccharide, iodine matter, Vitamin D and 10 various trace elements such as zinc, copper, manganese.
Adjuvant is the immunogenicity of one type of ability enhancement antigen, the material of raising vaccine effect, and the adjuvant technology makes homemade vaccine on price, have clear superiority just.In influenza vaccines, use the adjuvant not only can enhances immunogenicity, can also reduce the antigen consumption.When the reply pandemic influenza, the characteristic of using adjuvant to reduce antigen consumption and then expansion inoculation crowd quantity seems particularly important especially.The adjuvant that in the pandemic influenza vaccine research, is applied to mainly contains: aluminium adjuvant (aluminium oxide, aluminium hydroxide, aluminum phosphate), MF59 (are a kind of oil-in-water Emulsion; Comprise 1% zamene, 0.5%Tween80 and 0.5% 3 oleic acid gather the O/w emulsion of Pyrusussuriensis fat), LTK63 and novel adjuvant etc.
The adjuvant of listing influenza vaccines use mainly is aluminium adjuvant and MF59 at present.But a little less than in fact aluminium adjuvant induced ability that antibody produces to protein subunit vaccine, induce immune response also can produce the IgE relevant with allergy in some cases effectively; MF59 uses increasing the immunogenic while as adjuvant, has also strengthened reactionogenicity, symptom increases such as, pain red, swollen, hot like inoculation position.In addition, because problems such as safety, side effect, other materials such as LTK63 and novel adjuvant comprise that chitosan, ISCOMs etc. yet are only limited to conceptual phase as the influenza vaccines adjuvant.Therefore, along with various New Development influenzas in the outburst in the world with to the infective enhancing of people, accelerate the research of new type influenza vaccine, seek the needs that greater security and immunogenic adjuvant remain the influenza vaccines development.
It is owing to immunogenicity that can enhancement antigen, improve vaccine effect that adjuvant plays a role, so adjuvant generally can use in multiple vaccine, like present aluminium adjuvant.So the research of adjuvant is not only to influenza, and the control of other infectious disease also there is very high value.
Carapax Trionycis extract is the experimental result that example has obtained application as the application of immunological adjuvant with influenza (abbreviation influenza) viral vaccine.
Influenza (abbreviation influenza) is meant by influenza virus and causes to have the acute respiratory infectious disease of hyperinfection property.The influenza rapid onset, infectiousness is strong, and sickness rate is high, mainly through the air droplet transmission.Influenza popular do not have obvious seasonal, and just sickness rate is higher slightly at the beginning of last month of spring in winter, and almost all can take place every year; Perhaps local outburst, perhaps popular on a large scale, even cause being very popular of world scale; Serious harm human beings'health and life, caused white elephant to society.Up to the present, the inoculation influenza vaccines are considered to the best approach that flu-prevention takes place and propagates.Because the influenza virus variation is very fast; Usually annual popular type is all different; Manufacturing enterprise generally all selects the influenza virus strain to carry out production of vaccine under World Health Organization (WHO) (WHO) instructs, and the up-to-date influenza vaccines of annual inoculation just can reach preventive effect.In case it is extensive popular or worldwide popular that the influenza virus variation causes; The vaccine demand can roll up; But because the restriction of influenza vaccines production technology at present can not provide the capacity vaccine in the short time, so; Use immunological adjuvant, reduce the vaccine consumption and improve its effect and just seem extremely important.
Consult document and patent through system, do not find to have at present the report of immunoadjuvant function about Carapax Trionycis extract.
Summary of the invention
Single for solving present immunological adjuvant; Side effect is difficult to avoid; The production of vaccine supply is limited, and the vaccine demand is big when being difficult to deal with infectious disease and breaking out, and vaccine onset time is waited problem slowly; The present invention provides a kind of can improve immune effect of vaccine, brings the Carapax Trionycis immunological adjuvant of huge economic benefit and social benefit simultaneously.
Another object of the present invention is to provide a kind of influenza vaccines that contain the Carapax Trionycis immunological adjuvant.
Carapax Trionycis immunological adjuvant provided by the invention is made up of following proportion of masses:
Carapax Trionycis extract 0.001～100%
Said carrier is made up of following proportion of masses:
Emulsifying agent 20～70%
Co-emulsifier 20～70%
Oil phase 1～50%
Water 1～30%.
Said emulsifying agent is a kind of in conventional Tween80 or the polyoxyethylene castor oil;, be 1: 1 conventional polyoxyethylene castor oil and the mixture of Span80 perhaps perhaps for mass mixing ratio for mass mixing ratio is 3: 2～2: 3 conventional Tween80 and the mixture of Span80.
Said co-emulsifier is conventional 1, a kind of in 2-propylene glycol or the PEG400.
Said oil phase is a kind of in conventional isopropyl myristate or ethyl oleate or the injection soybean oil.
Said water is water for injection or the aqueous solution for injection that contains medicine or vaccine composition.
Carapax Trionycis immunological adjuvant provided by the invention, preferred following mass ratio:
Carapax Trionycis extract 0.1～100%
A kind of composition that contains the influenza vaccines of Carapax Trionycis immunological adjuvant provided by the invention is: the mass ratio of Carapax Trionycis immunological adjuvant and flu vaccine hemagglutinin is 50～180050, and wherein, said Carapax Trionycis immunological adjuvant is made up of following proportion of masses:
Carapax Trionycis extract 0.001～100%
Said Carapax Trionycis extract be with ethanol or water as solvent, extract through the conventional method of prior art and to obtain, specifically:
Ethanol extraction: exsiccant turtle ' s carapace powder is broken into powder, places in the container, extract with ethanol merceration or warm macerating, perhaps heating and refluxing extraction is filtered, and gets filtrating, reclaims the ethanol in the filtrating, and concentrated filtrate gets extractum; Wherein, when extracting, alcoholic acid mass concentration is 60～95% at every turn, and amount of alcohol added is 2～10 times of Carapax Trionycis quality of medicinal material, and merceration is at room temperature to soak 1～10 day, and warm macerating is under 40～60 ℃ of temperature, to soak 1～2 hour, and extraction time is 1～3 time; Heating and refluxing extraction is under 20～40 ℃ of temperature, reflux, extract, 1～3 time, each 20 minutes～4 hours;
Water extraction: exsiccant turtle ' s carapace powder is broken into powder, places in the container, extract with distilled water merceration or warm macerating, perhaps heating decocts and extracts, and filters, and gets filtrating, and concentrated filtrate gets extractum; Wherein, when extracting, the addition of distilled water is 2～10 times of Carapax Trionycis quality of medicinal material at every turn, and merceration is at room temperature to soak 1～10 day, and warm macerating is under 40～60 ℃ of temperature, to soak 1～2 hour, and extraction time is 1～3 time; Heating decocts to extract: each heated and boiled 20 minutes～4 hours, extraction time is 1～3 time.
In addition, said Carapax Trionycis extract also can adopt prior art except that water extraction, other conventional solvent extraction and the extract that obtains the ethanol extraction, like the extraction of adopting methanol, acetone, ethyl acetate etc. to carry out for solvent.
Carapax Trionycis immunological adjuvant provided by the invention is compared with traditional aluminum hydroxide adjuvant, has the following advantages:
1) simple, the economy of Carapax Trionycis extract method for preparing, environmental protection.
2) Carapax Trionycis is a kind of natural Chinese medicinal herb, and the Chang Zuowei health product use.In influenza vaccines, use as adjuvant, safe, side effect is little.For example with containing the influenza vaccines intramuscular injection mice of Carapax Trionycis extract, observe the mice activeness in 4 hours and do not have obviously and weaken, do not see systemic adverse reactions.And with the injection of aluminium hydroxide adjuvant influenza vaccines, can be observed the mice activeness obviously weakens, and injection site swelling is obvious.
3) use the influenza vaccines of Carapax Trionycis extract as adjuvant, method for preparing is easy, and vaccine quality is controlled easily; And be that adjuvant prepares influenza vaccines with the aluminium hydroxide, in preparation and the storing process, the influence factor is more, for difficult control of quality of vaccine.
4) with Carapax Trionycis extract as the influenza vaccines adjuvant, can better improve vaccine immunogenicity, reduce the vaccine antigen consumption greatly.For example through being the influenza vaccines of adjuvant with the Carapax Trionycis extract and being that the influenza vaccines immune mouse of adjuvant is found with the aluminium hydroxide; Even be the vaccine antigen consumption that uses of adjuvant with the Carapax Trionycis extract as the half the of the vaccine antigen consumption of adjuvant or still less, can reach ideal immune protective effect equally with aluminium hydroxide.
Description of drawings
Fig. 1 is the variation of tiring of the hemagglutination inhibition antibody of different vaccination group in 56 days;
The body weight gain situation that Fig. 2 is the different vaccination group in back 7 days of inoculation;
Fig. 3 tires for the hemagglutination inhibition antibody of different vaccination group in 42 days;
Fig. 4 tires at the hemagglutination inhibition antibody in 2 weeks, 4 weeks, 6 weeks for the different vaccination group.
The specific embodiment
Below in conjunction with embodiment the present invention is further specified, but do not limit the present invention with this.
The component that does not specialize in the embodiment of the invention is commercial product, and device therefor or instrument all are conventional.
Embodiment 1
The preparation of Carapax Trionycis ethanol extraction: get dry Carapax Trionycis 40g, be ground into powder, place in the container, use 4 times to be that the 160ml mass concentration is 65% ethanol; Carry out heating and refluxing extraction by routine, wherein, each heating-up temperature is 50 ℃; Return time is 60 minutes, and extraction time is 2 times, after the filtration; After filtrating carried out decompression recycling ethanol, remaining concentrating filter liquor was dry, extractum.
The preparation of Carapax Trionycis water extract: getting dry Carapax Trionycis 40g, be ground into powder, place in the container, is that the 320ml distilled water heats the decoction extraction by routine with 8 times; Wherein, each decocting time is 4 hours, and extraction time is 3 times, filters; Behind the merging filtrate, filtrating is carried out concentrate drying, get extractum.
Embodiment 3
The preparation of nano-emulsion carrier:
1) gets the raw materials ready by following mass ratio: 2g polyoxyethylene castor oil, 2g PEG400,2g isopropyl myristate, 1.07g water for injection;
2) in mixing blender jar, add the 2g polyoxyethylene castor oil, the 2g PEG400 is 300 rev/mins at rotating speed, under the room temperature condition, continues to stir 5 minutes, adds the 2g isopropyl myristate behind the mixing, and mix homogeneously gets mixture;
3) above-mentioned 2) mixture in splash into 1.07g water for injection, be 300 rev/mins simultaneously at rotating speed, under the room temperature, it is transparent stirring the mixture to liquid, obtains the nano-emulsion carrier;
4) to 3) the nano-emulsion carrier that makes detects as follows: 1. and detect through Ma Erwen particle diameter detector, particle size range is 28.706～46.952nm, and is qualified; 2. through 3000rpm, the 15min centrifugalize, the nano-emulsion system is not stratified, and is qualified; 3. warp was placed 50 days for 4 ℃, and nano-emulsion system no change is qualified.
Embodiment 4
1) takes by weighing the Carapax Trionycis extract 1.75mg of embodiment 2, be dissolved in the commercial H3N2 influenza virus cracking vaccine of 535ul (hemagglutinin content 43.67 μ g/ml), must contain the influenza vaccines of Carapax Trionycis extract;
2) get the 1g polyoxyethylene castor oil, the 1g PEG400, the 1g isopropyl myristate is put in the blender jar, and rotating speed maintains 300 rev/mins, and stirring at room 5 minutes to abundant mixing, gets mixing material;
3) above-mentioned 2) mixed solution in slowly splash into 1) preparation the influenza vaccines 535ul that contains Carapax Trionycis extract; Keep rotating speed simultaneously at 300 rev/mins; It is transparent being stirred to solution; Obtaining with Carapax Trionycis extract, nano-emulsion carrier is the influenza virus vaccine of adjuvant, and wherein: the mass ratio of Carapax Trionycis extract is 0.058%, and the carrier mass ratio is 99.942%; The mass ratio of Carapax Trionycis immunological adjuvant and flu vaccine hemagglutinin is: 128500.
4) to 3) influenza virus vaccine that contains the Carapax Trionycis immunological adjuvant that makes detects as follows: 1. and detect through Ma Erwen particle diameter detector, particle size range is 32.724～46.435nm, and is qualified; 2. through 3000rpm, the 15min centrifugalize, the nano-emulsion system is not stratified, and is qualified; 3. warp was placed 50 days for 4 ℃, and nano-emulsion system no change is qualified.
With the influenza virus vaccine that contains the Carapax Trionycis immunological adjuvant of embodiment 4 preparations, injection standard body weight Male Kunming strain mice, every mice 200ul; Flu vaccine hemagglutinin content is 1.3ug in this 200ul vaccinate, Carapax Trionycis 0.1mg, the 1st day inoculation one pin; Reseeded a pin on the 21st day, first week beginning after first immunisation, every tail blood of adopting at a distance from 7 days; Separation of serum is measured blood clotting and is suppressed to tire, set up simultaneously injection equivalent normal saline blank group, etc. the Al (OH) of hemagglutinin content
3The adjuvant matched group of adjuvant influenza vaccines, etc. the independent influenza vaccines matched group of hemagglutinin content.The hemagglutination inhibition antibody that the obtains testing result of tiring is seen accompanying drawing 1.
Embodiment 5
1) takes by weighing the Carapax Trionycis extract 3.5mg of embodiment 1, be dissolved in the commercial H3N2 influenza virus cracking vaccine of 382ul (hemagglutinin content is 43.67 μ g/ml), must contain the influenza vaccines of Carapax Trionycis extract;
2) take by weighing the 0.5g polyoxyethylene castor oil, 0.5g Span80,1g 1, and 2-propylene glycol, 1g isopropyl myristate are in blender jar, and rotating speed maintains 200 rev/mins, and stirring at room 5 minutes to abundant mixing, gets mixed solution;
3) above-mentioned 2) mixed solution in slowly splash into 1) preparation the influenza vaccines 400ul that contains Carapax Trionycis extract; Keep rotating speed simultaneously at 100 rev/mins; Being stirred to solution is that yellowish-brown is transparent, and obtaining with Carapax Trionycis extract, nano-emulsion carrier is the influenza virus vaccine of adjuvant.Wherein: the mass ratio of Carapax Trionycis extract is 0.117%, and the carrier mass ratio is 99.883%; The mass ratio of Carapax Trionycis immunological adjuvant and flu vaccine hemagglutinin is: 180045.
Embodiment 6
1) following different adjuvant of preparation or solution:
A) the Carapax Trionycis extract 0.2mg that gets embodiment 2 is dissolved in 160 μ l normal saline, and is subsequent use;
B) get the aluminium hydroxide 12.5 μ l that concentration is 15.98mg/ml, wherein contain AL (OH)
30.2mg, add normal saline to volume and be 160 μ l, subsequent use;
C) 160 μ l normal saline are subsequent use;
D) 200 μ l normal saline are subsequent use;
2) the following different vaccine of preparation:
Getting hemagglutinin content is the commercial H3N2 influenza virus cracking vaccine 40 μ l of 43.67 μ g/ml, mixes with above-mentioned a, b, c different solutions respectively, obtain respectively containing the Carapax Trionycis immunological adjuvant influenza vaccines sample A, contain AL (OH)
3The influenza vaccines sample B of adjuvant, independent influenza vaccines sample C.The d group does not add vaccine, is normal saline blank sample D.The above-mentioned A that is disposed, B, C, four samples of D all prepare three parts, and volume is 200 μ l, and is subsequent use; Wherein contain among the influenza vaccines sample A of Carapax Trionycis immunological adjuvant: the mass ratio of Carapax Trionycis extract is 100%, and the carrier mass ratio is 0%; The mass ratio of Carapax Trionycis immunological adjuvant and flu vaccine hemagglutinin is 144.
3) immunity inoculation and detection:
With above-mentioned 2) four different samples of the A, B, C, the D that are joined female kunming mices of immune SBW respectively, 3 every group, every intramuscular injection 200ul, the 1st day inoculation one pin reseeded a pin on the 21st day.Before inoculation in first day, each was organized mice in the 8th day with inoculation back and carry out measured body weight, the body weight of respectively organizing balanced growth in 7 days is calculated, the result sees accompanying drawing 2; And after first immunisation, got tail blood in 14 days, 21 days, 28 days, 35 days, 42 days, and separation of serum is measured blood clotting and is suppressed to tire, and the result sees accompanying drawing 3.
Embodiment 7
1) the following different vaccine of preparation:
A, the Carapax Trionycis extract 0.3mg that gets embodiment 1 are dissolved in 178.5 μ l normal saline; Add then commercial H3N2 influenza split vaccine 20 μ l to final volume be 200 μ l; The hemagglutinin content of H3N2 influenza split vaccine is 0.87ug, must contain the influenza virus vaccine of Carapax Trionycis immunological adjuvant, and is subsequent use; Wherein: the mass ratio of Carapax Trionycis extract is 100%, and the carrier mass ratio is 0%; The mass ratio of Carapax Trionycis immunological adjuvant and flu vaccine hemagglutinin is: 345.
B, the Carapax Trionycis extract 0.1mg that gets embodiment 2 are dissolved in the 160 μ l normal saline; Add commercial H3N2 influenza split vaccine 40 μ l then to final volume 200 μ l; The hemagglutinin content of H3N2 influenza split vaccine is 1.75 μ g, must contain the influenza virus vaccine of Carapax Trionycis immunological adjuvant, and is subsequent use; Wherein: the mass ratio of Carapax Trionycis extract is 100%, and the carrier mass ratio is 0%; The mass ratio of Carapax Trionycis immunological adjuvant and flu vaccine hemagglutinin is: 57.
C, get H3N2 influenza split vaccine 40 μ l, wherein contain hemagglutinin 1.75 μ g, being mixed to final volume with 160 μ l normal saline is 200 μ l, as independent vaccine control group, subsequent use;
D, get the aluminum hydroxide adjuvant 37.5 μ l that concentration is 15.98mg/ml, wherein contain 0.6mgAL (OH)
3, 40 μ l mix with the influenza vaccines that contain hemagglutinin 1.75 μ g, add normal saline to final volume 200 μ l then, as AL (OH)
3The Adjuvanted vaccines matched group, subsequent use:
E, normal saline 200 μ l, for the blank group, subsequent use;
2) immunity inoculation and antibody test:
With the above-mentioned A for preparing, B, C, D, five immune SBWs of samples difference of E Kunming female mice, intramuscular injection, every 200 μ l; Every group 5, the 1st day inoculation one pin reseeded a pin on the 21st day; Respectively at carrying out the afterbody blood sampling in 2 weeks, 4 weeks, 6 weeks after the first immunisation; Separation of serum detects hemagglutination inhibition antibody and tires, and the result sees accompanying drawing 4.
Conclusion: Carapax Trionycis immunological adjuvant provided by the invention has significant immunological adjuvant effect, and toxic and side effects is little, can be used for the multiple vaccine of people, animal, to prevent and treat various infectious disease.
Claims (3)
1. the Carapax Trionycis immunological adjuvant of influenza vaccines is characterized in that being made up of following proportion of masses:
Carapax Trionycis extract 0.001～100%
Carrier 0～99.999%
Said Carapax Trionycis extract is the ethanol extraction of Carapax Trionycis or the water extract of Carapax Trionycis, wherein:
The ethanol extraction of Carapax Trionycis prepares through following method: get dry Carapax Trionycis 40g, be ground into powder, place in the container, use 4 times to be that the 160ml mass concentration is 65% ethanol; Carry out heating and refluxing extraction by routine, wherein, each heating-up temperature is 50 ℃; Return time is 60 minutes, and extraction time is 2 times, after the filtration; After filtrating carried out decompression recycling ethanol, remaining concentrating filter liquor was dry, extractum;
The water extract of Carapax Trionycis prepares through following method: get dry Carapax Trionycis 40g, be ground into powder, place in the container; With 8 times is that the 320ml distilled water heats the decoction extraction by routine, and wherein, each decocting time is 4 hours; Extraction time is 3 times, filters, behind the merging filtrate; Filtrating is carried out concentrate drying, get extractum;
Being prepared as of said carrier:
In mixing blender jar, add the 2g polyoxyethylene castor oil, the 2g PEG400 is 300 rev/mins at rotating speed, under the room temperature condition, continues to stir 5 minutes, adds the 2g isopropyl myristate behind the mixing, and mix homogeneously gets mixture; In said mixture, splashing into 1.07g water for injection, is 300 rev/mins at rotating speed simultaneously, and under the room temperature, it is transparent stirring the mixture to liquid, obtains the nano-emulsion carrier;
Perhaps
Get the 1g polyoxyethylene castor oil, the 1g PEG400, the 1g isopropyl myristate is put in the blender jar, and rotating speed maintains 300 rev/mins, and stirring at room 5 minutes to abundant mixing, gets mixing material;
Perhaps
Take by weighing the 0.5g polyoxyethylene castor oil, 0.5g Span80,1g 1, and 2-propylene glycol, 1g isopropyl myristate are in blender jar, and rotating speed maintains 200 rev/mins, and stirring at room 5 minutes to abundant mixing, gets mixed solution.
2. the Carapax Trionycis immunological adjuvant of influenza vaccines as claimed in claim 1 is characterized in that being made up of following proportion of masses:
Carapax Trionycis extract 0.1～100%
Carrier 0～99.9%.
3. influenza vaccines that contain the Carapax Trionycis immunological adjuvant, the mass ratio that it is characterized in that Carapax Trionycis immunological adjuvant and flu vaccine hemagglutinin is 50～180050, wherein, said Carapax Trionycis immunological adjuvant is made up of following proportion of masses:
Carapax Trionycis extract 0.001～100%
Carrier 0～99.999%
Said Carapax Trionycis extract is the ethanol extraction of Carapax Trionycis or the water extract of Carapax Trionycis, wherein:
The ethanol extraction of Carapax Trionycis prepares through following method: get dry Carapax Trionycis 40g, be ground into powder, place in the container, use 4 times to be that the 160ml mass concentration is 65% ethanol; Carry out heating and refluxing extraction by routine, wherein, each heating-up temperature is 50 ℃; Return time is 60 minutes, and extraction time is 2 times, after the filtration; After filtrating carried out decompression recycling ethanol, remaining concentrating filter liquor was dry, extractum;
The water extract of Carapax Trionycis prepares through following method: get dry Carapax Trionycis 40g, be ground into powder, place in the container; With 8 times is that the 320ml distilled water heats the decoction extraction by routine, and wherein, each decocting time is 4 hours; Extraction time is 3 times, filters, behind the merging filtrate; Filtrating is carried out concentrate drying, get extractum;
Being prepared as of said carrier:
In mixing blender jar, add the 2g polyoxyethylene castor oil, the 2g PEG400 is 300 rev/mins at rotating speed, under the room temperature condition, continues to stir 5 minutes, adds the 2g isopropyl myristate behind the mixing, and mix homogeneously gets mixture; In said mixture, splashing into 1.07g water for injection, is 300 rev/mins at rotating speed simultaneously, and under the room temperature, it is transparent stirring the mixture to liquid, obtains the nano-emulsion carrier;
Perhaps
Get the 1g polyoxyethylene castor oil, the 1g PEG400, the 1g isopropyl myristate is put in the blender jar, and rotating speed maintains 300 rev/mins, and stirring at room 5 minutes to abundant mixing, gets mixing material;
Perhaps
Take by weighing the 0.5g polyoxyethylene castor oil, 0.5g Span80,1g 1, and 2-propylene glycol, 1g isopropyl myristate are in blender jar, and rotating speed maintains 200 rev/mins, and stirring at room 5 minutes to abundant mixing, gets mixed solution.
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Non-Patent Citations (3)
| Title |
|---|
| 于力等.纳米乳的研究及其在制剂学领域的应用.《药学进展》.2006,第30卷(第11期),491-497. * |
| 李璐璐等.中药免疫增强剂在动物生产中的应用.《兽药研究与应用》.2009,(第3期),32-35. * |
| 陈士奎主编.中西医结合药理学研究进展.《中西医结合医学导论》.中国中医药出版社,2005,170. * |
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