CN104383534A - Immunological adjuvant containing fucosan sulfate and application of immunological adjuvant - Google Patents
Immunological adjuvant containing fucosan sulfate and application of immunological adjuvant Download PDFInfo
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- CN104383534A CN104383534A CN201410579310.3A CN201410579310A CN104383534A CN 104383534 A CN104383534 A CN 104383534A CN 201410579310 A CN201410579310 A CN 201410579310A CN 104383534 A CN104383534 A CN 104383534A
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Abstract
The invention relates to an immunological adjuvant containing fucosan sulfate and application of the immunological adjuvant. The technical problems that at present, the kinds of immunological adjuvants are few, side effects cannot be avoided, and a great number of vaccines need to be provided when infectious diseases break out are solved. The immunological adjuvant consists of the following components in mass ratio: fucosan sulfate, ginsenoside and the rest of carriers, wherein the carriers consist of the following components in mass ratio: an emulsifying agent, an emulsifying auxiliary agent, oil phase and aqueous phase. The immunological adjuvant can be widely used in the preparation field of vaccines.
Description
Technical field
The present invention relates to a kind of immunological adjuvant containing fucoidan, also relate to the influenza vaccines of the immunological adjuvant containing fucoidan, belong to biotechnology and biomedicine field.
Background technology
Fucoidan is as the intrinsic iuntercellular polysaccharide of the Brown algaes such as Thallus Laminariae (Thallus Eckloniae), and being present in Brown algae cell wall matrix, is bioactive substance specific to Brown algae.Such as, the fucoidan (Fucoidan) extracted for raw material with Thallus Laminariae (Thallus Eckloniae) (Laminariajaponicia), forms primarily of sulfur acid base fucose (Fucose); Secondly, also with a small amount of galactose, mannose, xylose, arabinose, alduronic acid etc., be a kind of water miscible assorted poly-polysaccharide, the fucoidan that is otherwise known as, sulfated fucan, it mainly contains effective constituent is α-L-fucose-4-sulfuric ester.
Fucoidan is a kind of outstanding health food, pharmaceutical raw material, because of natural containing sulfate radical, thus has the characteristic of anionic polymer compound.Research shows, fucoidan has anticoagulation, blood fat reducing, anti-chronic kidney hypofunction, antitumor, antiviral, promotion tissue regeneration, suppresses the multiple physiologically active such as gastric ulcer, enhancing human body immunity function.Fucoidan has the pharmacological activity of obvious anticoagulation and fibrinolysis enhancing; Comprise fucose or its degradation product energy cancer cell specific induction of apoptosis of sulfate, can be used as inducer of apoptosis and anticarcinogen; The generation of cell growth factor can be brought out, thus promote various Growth of Cells, repair and be damaged or hypothyroid Organ and tissue; Because it has anticoagulant effect, so be applicable to the high patient of blood viscosity, can be used as medicine or the health product of antithrombotic; It also has effect of good blood fat reducing, blood sugar lowering, cholesterol reducing, can overcome some side effect of fat-reducing medicament; Can chronic kidney hypofunction being treated, the early stage renal failure Be very effective of centering, especially to improving renal function, improving kidney particularly remarkable to creatinine clearance rate effect; Can be used as bonding agent and the resistance vapor of metal ion, as human body can be made, more than 70% is reduced to the absorption of lead.
In addition, fucoidan can not only effective immune cell activated, improves immunity of organisms, and to the toxic and side effects of normal host cell without teratogenesis, is therefore considered to a kind of important biological regulator.In addition, Ginsenosides Rg1 and Rb1 can improve immunity of organisms, and they can be used as immunological adjuvant bibliographical information, the immunogenicity of enhancement antigen.
Adjuvant mainly aluminium adjuvant and the MF59 of influenza vaccines use in the market, but in fact the ability that produces protein subunit vaccine induction of antibodies of aluminium adjuvant is more weak, can not induce immune response effectively, also can produce the IgE relevant with anaphylaxis in some cases, its side effect cannot be avoided; When MF59 is as vaccine adjuvant, the while of increasing immunogenic, too increase reactionogenicity, as symptoms such as inoculation position red and swollen heat pains.Along with various new influenza is in global outburst with to the infective enhancing of people, in the urgent need to exploitation, there is greater security and immunogenic immunological adjuvant.
Summary of the invention
Just in order to solve, current immunological adjuvant kind is few, side effect cannot be avoided in the present invention, can not need to provide the technical problem of a large amount of vaccine when breaking out by gratification infectious diseases, provides a kind of immunological adjuvant containing fucoidan that effectively can improve immune effect of vaccine, few side effects, reduction vaccine consumption.
For this reason, the invention provides a kind of immunological adjuvant containing fucoidan, it is made up of the component of following masses ratio: fucoidan 1 ~ 20%; Ginsengenin 20 .01 ~ 2%; Remaining is carrier; Carrier is made up of the component of following masses ratio: emulsifying agent 1 ~ 30%; Co-emulsifier 1 ~ 50%; Oil phase 1 ~ 50%; Remaining is aqueous phase.
Preferably, ginsenoside is the one in Rg1 or Rb1, or for mass mixing thing is than the mixture of Rg1 and the Rb1 for 1:100 ~ 100:1.
Preferably, emulsifying agent is the one in conventional Tween 80 or polyoxyethylene castor oil, or for mass mixing ratio is the conventional Tween 80 of 3:1 ~ 1:3 and the mixture of Span80, or for mass mixing ratio be the conventional polyoxyethylene castor oil of 1:1 and the mixture of Span80.
Preferably, co-emulsifier is the one in conventional 1,2-PD or PEG400.
Preferably, oil phase is the one in conventional isopropyl myristate or ethyl oleate or injection soybean oil.
Preferably, aqueous phase is water for injection or the aqueous solution for injection containing medicine or vaccine composition.
Invention also provides the application of immunological adjuvant in preparation influenza vaccines containing fucoidan, immunological adjuvant containing fucoidan and the mass ratio of flu vaccine hemagglutinin are 6000 ~ 321000:1, wherein, the immunological adjuvant containing fucoidan is made up of the component of following masses ratio: fucoidan 1 ~ 20%; Ginsengenin 20 .01 ~ 2%; Remaining is carrier.
The present invention contains the immunological adjuvant of fucoidan and ginsenoside, can the immunogenicity of remarkable enhancement antigen, improves the effect of vaccine, makes vaccine in price, have obvious advantage.Especially in influenza vaccines, use adjuvant, not only can strengthen immunogenicity, antigen consumption can also be reduced, especially when tackling flu outbreak, the use of adjuvant can effectively reduce antigen consumption, expands inoculation crowd quantity, reduces the pressure that production of vaccine enterprise brings because of underproduce.
Detailed description of the invention
In order to more clearly understand technology contents of the present invention, the special following examples that adopt are described in detail, and its object is to better understand content of the present invention but not limit the scope of the invention.
In the following example of the present invention, ginsenoside Rg1 or Rb1 are commercial products or extract the product obtained for conventional method, and product purity is more than 95%.Such as, Nanjing is for the ginsenoside Rb1 of this Chinese mugwort traditional Chinese medicine technology institute, and production code member TCM-039, HPLC detect purity and be not less than 98%; Ginsenoside Rg1, production code member TCM-041, HPLC detect purity and are not less than 98%.
The product that fucoidan is commercial products or obtains for conventional method, product purity is more than 80%, and all the other 20% are other polysaccharide component.Such as, the product of sigma company, purity is not less than 85%, production code member F8065.
H1N1 influenza virus cracking vaccine, such as, the influenza A H1N1 influenza virus split vaccine that Zhejiang Tianyuan Biological Pharmaceutical Co., Ltd. produces, the accurate word S20090027 of traditional Chinese medicines; The influenza A H1N1 influenza virus split vaccine that the effective company of the blue biovaccine of China produces, the accurate word S20090016 of traditional Chinese medicines.
Embodiment 1
The preparation (A) of nano-emulsion carrier:
(1) take 1.5g Tween 80 respectively, 1.0g Span80,1.5g molecular weight is 400 daltonian Polyethylene Glycol, join in glass container, stirred at ambient temperature 3 ~ 10 minutes, rotating speed 300 revs/min, add 4g injection soybean oil after mixing, continue to be uniformly mixed and obtain mixture;
(2) in the mixture of above-mentioned steps (1), instill 4g water for injection, room temperature, stir 5 ~ 15 minutes under 260 revs/min, obtain nano-emulsion carrier.
Embodiment 2
The preparation (B) of nano-emulsion carrier:
(1) 0.04g Tween 80 is taken respectively, 0.04g Span80,0.12g molecular weight is 400 daltonian Polyethylene Glycol, join in glass container, stirred at ambient temperature 3 ~ 10 minutes, rotating speed 300 revs/min, adds 4g injection soybean oil after mixing, continue to be uniformly mixed to obtain mixture;
(2) in the mixture of above-mentioned steps (1), instill 4g water for injection, room temperature, stir 5 ~ 15 minutes under 260 revs/min, obtain nano-emulsion carrier.
Embodiment 3
The preparation (C) of nano-emulsion carrier:
(1) 0.4g Tween 80 is taken respectively, 0.8g Span80,1.2g molecular weight is 400 daltonian Polyethylene Glycol, join in glass container, stirred at ambient temperature 3 ~ 10 minutes, rotating speed 300 revs/min, adds 0.05g injection soybean oil after mixing, continue to be uniformly mixed to obtain mixture;
(2) in the mixture of above-mentioned steps (1), instill 4.0g water for injection, room temperature, stir 5 ~ 15 minutes under 260 revs/min, obtain nano-emulsion carrier.
Embodiment 4
(1) take commercial fucoidan 162mg, 1.62mg ginsenoside Rg1 is dissolved in 1.0ml water, is mixed with aqueous solution;
(2) aqueous solution above-mentioned (1) prepared and 4.0ml city buy H1N1 influenza virus cracking vaccine (hemagglutinin content is 30 μ g/ml) and mix, and then 1.0ml normal saline is added in its mixture, obtain the influenza vaccines containing fucoidan and ginsenoside Rg1 that cumulative volume is 6ml, in temperature 4 DEG C preservation, for subsequent use;
(3) take 6g injection soybean oil, the daltonian Polyethylene Glycol of 2.0g molecular weight 400,0.5g Tween 80,1.5g Span80 is in glass container, and temperature 4 DEG C, stirs 5-10 minute, fully mix, obtain mixed solution under 220 revs/min;
(4) in the mixed solution of above-mentioned (3), slowly instill the influenza vaccines 6ml containing fucoidan and ginsenoside Rg1 of (2), temperature 4 DEG C, under 220 revs/min, stir 5-30 minute, obtaining containing fucoidan, nano-emulsion is the influenza virus vaccine of adjuvant.
Embodiment 5
(1) take commercial fucoidan 1.2g, 10mg ginsenoside Rb1 is dissolved in 2ml water, is mixed with aqueous solution;
(2) aqueous solution above-mentioned (1) prepared and 2.2ml city buy H1N1 influenza virus cracking vaccine (hemagglutinin content is 30 μ g/ml) and mix, and then 1.8ml normal saline is added in its mixture, obtain the influenza vaccines containing fucoidan and ginsenoside Rg1 that cumulative volume is 6ml, in temperature 4 DEG C preservation, for subsequent use;
(3) take 5.8g injection soybean oil, the daltonian Polyethylene Glycol of 2.2g molecular weight 400,0.6 Tween 80,1.4g Span80 is in glass container, and temperature 4 DEG C, stirs 5-10 minute, fully mix, obtain mixed solution under 220 revs/min;
(4) in the mixed solution of above-mentioned (3), slowly instill the influenza vaccines 6ml containing fucoidan and ginsenoside Rb1 of (2), temperature 4 DEG C, under 220 revs/min, stir 5-30 minute, obtaining containing fucoidan, nano-emulsion is the influenza virus vaccine of adjuvant.
Embodiment 6
(1) take commercial fucoidan 4.0g, 5mg ginsenoside Rg1,5mg ginsenoside Rb1, be dissolved in 2ml water, be mixed with aqueous solution;
(2) aqueous solution above-mentioned (1) prepared and 2.1ml city buy H1N1 influenza virus cracking vaccine (hemagglutinin content is 30 μ g/ml) and mix, and then 1.9ml normal saline is added in its mixture, obtain the influenza vaccines containing fucoidan and ginsenoside Rg1 that cumulative volume is 6ml, in temperature 4 DEG C preservation, for subsequent use;
(3) take 6.5g injection soybean oil, the daltonian Polyethylene Glycol of 1.5g molecular weight 400,1.2 Tween 80s, 0.8g Span80 is in glass container, and temperature 4 DEG C, stirs 5-10 minute, fully mix, obtain mixed solution under 260 revs/min;
(4) in the mixed solution of above-mentioned (3), slowly instill the influenza vaccines 6ml containing fucoidan and ginsenoside Rg1 and Rb1 of (2), temperature 4 DEG C, under 220 revs/min, stir 5-30 minute, obtaining containing fucoidan, nano-emulsion is the influenza virus vaccine of adjuvant.
Embodiment 7
(1) take commercial fucoidan 1.0g, 0.34g ginsenoside Rb1 is dissolved in 2ml water, is mixed with aqueous solution;
(2) aqueous solution above-mentioned (1) prepared and 2.5ml city buy H1N1 influenza virus cracking vaccine (hemagglutinin content is 30 μ g/ml) and mix, and then 1.5ml normal saline is added in its mixture, obtain the influenza vaccines containing fucoidan and ginsenoside Rg1 that cumulative volume is 6ml, in temperature 4 DEG C preservation, for subsequent use;
(3) 6.1g injection soybean oil is taken, the daltonian Polyethylene Glycol of 1.9g molecular weight 400,1.0 polyoxyethylene castor oils, 1.0g Span80 is in glass container, and temperature 4 DEG C, stirs 5-10 minute under 230 revs/min, abundant mixing, obtains mixed solution;
(4) in the mixed solution of above-mentioned (3), slowly instill the influenza vaccines 6ml containing fucoidan and ginsenoside Rb1 of (2), temperature 4 DEG C, under 220 revs/min, stir 5-30 minute, obtaining containing fucoidan, nano-emulsion is the influenza virus vaccine of adjuvant.
Embodiment 8
(1) take commercial fucoidan 200mg, 20mg ginsenoside Rg1 is dissolved in 2.1ml water, is mixed with aqueous solution;
(2) aqueous solution above-mentioned (1) prepared and 2.0ml city buy H1N1 influenza virus cracking vaccine (hemagglutinin content is 30 μ g/ml) and mix, and then 1.9ml normal saline is added in its mixture, obtain the influenza vaccines containing fucoidan and ginsenoside Rg1 that cumulative volume is 6.0ml, in temperature 4 DEG C preservation, for subsequent use;
(3) take 6.3g injection soybean oil, the daltonian Polyethylene Glycol of 1.7g molecular weight 400,1.3 Tween 80s, 0.7g Span80 is in glass container, and temperature 4 DEG C, stirs 5-10 minute, fully mix, obtain mixed solution under 210 revs/min;
(4) in the mixed solution of above-mentioned (3), slowly instill the influenza vaccines 6.3ml containing fucoidan and ginsenoside Rg1 of (2), temperature 4 DEG C, under 220 revs/min, stir 5-30 minute, obtaining containing fucoidan, nano-emulsion is the influenza virus vaccine of adjuvant.
Embodiment 9
(1) take commercial fucoidan 300mg, 30mg ginsenoside Rb1 is dissolved in 1.8ml water, is mixed with aqueous solution;
(2) aqueous solution above-mentioned (1) prepared and 1.7ml city buy H1N1 influenza virus cracking vaccine (hemagglutinin content is 30 μ g/ml) and mix, and then 2.5ml normal saline is added in its mixture, obtain the influenza vaccines containing fucoidan and ginsenoside Rg1 that cumulative volume is 6.0ml, in temperature 4 DEG C preservation, for subsequent use;
(3) take 5.5g injection soybean oil, the daltonian Polyethylene Glycol of 2.5g molecular weight 400,0.9 Tween 80,1.1g Span80 is in glass container, and temperature 4 DEG C, stirs 5-10 minute, fully mix, obtain mixed solution under 210 revs/min;
(4) in the mixed solution of above-mentioned (3), slowly instill the influenza vaccines 6.0ml containing fucoidan and ginsenoside Rb1 of (2), temperature 4 DEG C, under 220 revs/min, stir 5-30 minute, obtaining containing fucoidan, nano-emulsion is the influenza virus vaccine of adjuvant.
Embodiment 10
(1) take commercial fucoidan 0.150g, 1.5mg ginsenoside Rg1 is dissolved in 1.0ml water, is mixed with aqueous solution;
(2) H1N1 influenza virus cracking vaccine being concentrated into hemagglutinin content is 0.2mg/ml, then the influenza virus cracking vaccine after being concentrated with 10ml by the aqueous solution that above-mentioned (1) prepares mixes, obtain the influenza vaccines containing fucoidan and ginsenoside Rg1 that cumulative volume is 11ml, in temperature 4 DEG C preservation, for subsequent use;
(3) take 0.12g injection soybean oil, the daltonian Polyethylene Glycol of 0.12g molecular weight 400,0.6g Tween 80,0.6g Span80 is in glass container, and temperature 4 DEG C, stirs 5-10 minute, fully mix, obtain mixed solution under 220 revs/min;
(4) in the mixed solution of above-mentioned (3), slowly instill the influenza vaccines 11ml containing fucoidan and ginsenoside Rg1 of (2), temperature 4 DEG C, under 220 revs/min, stir 5-30 minute, obtaining containing fucoidan, nano-emulsion is the influenza virus vaccine of adjuvant.
Claims (7)
1. the immunological adjuvant containing fucoidan, is characterized in that being made up of the component of following masses ratio: fucoidan 1 ~ 20%; Ginsengenin 20 .01 ~ 2%; Remaining is carrier;
Described carrier is made up of the component of following masses ratio: emulsifying agent 1 ~ 30%; Co-emulsifier 1 ~ 50%; Oil phase 1 ~ 50%; Remaining is aqueous phase.
2. the immunological adjuvant containing fucoidan according to claim 1, it is characterized in that, described ginsenoside is the one in Rg1 or Rb1, or for mass mixing thing is than the mixture of Rg1 and the Rb1 for 1:100 ~ 100:1.
3. the immunological adjuvant containing fucoidan according to claim 1, it is characterized in that, described emulsifying agent is the one in conventional Tween 80 or polyoxyethylene castor oil, or for mass mixing ratio is the conventional Tween 80 of 3:1 ~ 1:3 and the mixture of Span80, or for mass mixing ratio be the conventional polyoxyethylene castor oil of 1:1 and the mixture of Span80.
4. the immunological adjuvant containing fucoidan according to claim 1, is characterized in that, described co-emulsifier is the one in conventional 1,2-PD or PEG400.
5. the immunological adjuvant containing fucoidan according to claim 1, is characterized in that, described oil phase is the one in conventional isopropyl myristate or ethyl oleate or injection soybean oil.
6. the immunological adjuvant containing fucoidan according to claim 1, is characterized in that, described aqueous phase is water for injection or the aqueous solution for injection containing medicine or vaccine composition.
7. the application of the immunological adjuvant as claimed in claim 1 containing fucoidan in preparation influenza vaccines, it is characterized in that, immunological adjuvant containing fucoidan and the mass ratio of flu vaccine hemagglutinin are 6000 ~ 321000:1, wherein, the described immunological adjuvant containing fucoidan is made up of the component of following masses ratio: fucoidan 1 ~ 20%; Ginsengenin 20 .01 ~ 2%; Remaining is carrier.
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Cited By (6)
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| CN105816873A (en) * | 2016-03-11 | 2016-08-03 | 中牧实业股份有限公司 | Polymer combined adjuvant of animal vaccine and applications thereof |
| CN108210922A (en) * | 2018-01-18 | 2018-06-29 | 浙江普康生物技术股份有限公司 | A kind of novel cellular immunity enhancing adjuvant |
| RU2676266C2 (en) * | 2017-06-14 | 2018-12-27 | Федеральное государственное бюджетное научное учреждение "Научно-исследовательский институт эпидемиологии и микробиологии имени Г.П. Сомова" (НИИ эпидемиологии и микробиологии имени Г.П. Сомова") | Adjuvant for anti-viral vaccines |
| CN112294955A (en) * | 2020-10-28 | 2021-02-02 | 国药中生生物技术研究院有限公司 | Compound immunologic adjuvant and application thereof |
| CN112546056A (en) * | 2020-12-11 | 2021-03-26 | 南京医科大学 | Composition for treating chemotherapy-induced peripheral neuropathy and application thereof |
| CN114917334A (en) * | 2022-07-21 | 2022-08-19 | 天津三江永利生物科技有限公司 | Water-in-oil-in-water type composite vaccine adjuvant, and preparation method and application thereof |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105816873A (en) * | 2016-03-11 | 2016-08-03 | 中牧实业股份有限公司 | Polymer combined adjuvant of animal vaccine and applications thereof |
| CN105816873B (en) * | 2016-03-11 | 2019-07-26 | 中牧实业股份有限公司 | A kind of combination of polymers adjuvant of animal vaccine and its application |
| RU2676266C2 (en) * | 2017-06-14 | 2018-12-27 | Федеральное государственное бюджетное научное учреждение "Научно-исследовательский институт эпидемиологии и микробиологии имени Г.П. Сомова" (НИИ эпидемиологии и микробиологии имени Г.П. Сомова") | Adjuvant for anti-viral vaccines |
| CN108210922A (en) * | 2018-01-18 | 2018-06-29 | 浙江普康生物技术股份有限公司 | A kind of novel cellular immunity enhancing adjuvant |
| CN112294955A (en) * | 2020-10-28 | 2021-02-02 | 国药中生生物技术研究院有限公司 | Compound immunologic adjuvant and application thereof |
| CN112546056A (en) * | 2020-12-11 | 2021-03-26 | 南京医科大学 | Composition for treating chemotherapy-induced peripheral neuropathy and application thereof |
| CN114010653A (en) * | 2020-12-11 | 2022-02-08 | 南京医科大学 | Application of composition containing ginsenoside Rg1 and fucoidin in preparation of medicine for reducing expression of tissue factor |
| CN114028416A (en) * | 2020-12-11 | 2022-02-11 | 南京医科大学 | Composition for treating chemotherapy-induced peripheral neuropathy and application thereof |
| CN112546056B (en) * | 2020-12-11 | 2022-02-25 | 南京医科大学 | Composition for treating chemotherapy-induced peripheral neuropathy and application thereof |
| CN114028416B (en) * | 2020-12-11 | 2023-07-25 | 南京医科大学 | Composition for treating chemotherapy-induced peripheral neuropathy and application thereof |
| CN114010653B (en) * | 2020-12-11 | 2023-07-25 | 南京医科大学 | Application of composition comprising ginsenoside Rg1 and fucoidin in preparation of medicine for reducing expression of tissue factor |
| CN114917334A (en) * | 2022-07-21 | 2022-08-19 | 天津三江永利生物科技有限公司 | Water-in-oil-in-water type composite vaccine adjuvant, and preparation method and application thereof |
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