CN110507819B - Application of artesunate as immunologic adjuvant in preparation of rabies vaccine - Google Patents
Application of artesunate as immunologic adjuvant in preparation of rabies vaccine Download PDFInfo
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- CN110507819B CN110507819B CN201910745298.1A CN201910745298A CN110507819B CN 110507819 B CN110507819 B CN 110507819B CN 201910745298 A CN201910745298 A CN 201910745298A CN 110507819 B CN110507819 B CN 110507819B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/20011—Rhabdoviridae
- C12N2760/20111—Lyssavirus, e.g. rabies virus
- C12N2760/20134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Abstract
The invention provides application of artesunate as an immunologic adjuvant in preparation of rabies vaccines, and provides an Artesunate (ART) artemisinin derivative as a rabies vaccine adjuvant for the first time. The invention discovers that the Artesunate (ART) serving as the artemisinin derivative can obviously enhance the immunogenicity of the Artesunate (ART) serving as a rabies vaccine adjuvant and can serve as a novel rabies vaccine candidate adjuvant through experimental research. The research on the effect of the vaccine shows that Artesunate (ART) with lower dose can promote the immune effect of rabies vaccine and can reduce the cost for preparing the vaccine. The vaccine effect research of the invention shows that the Artesunate (ART) which is an artemisinin derivative can generate better protective force as a rabies vaccine adjuvant.
Description
Technical Field
The invention belongs to the field of molecular biological immunology. More particularly, relates to application of artesunate as an immunologic adjuvant in preparation of rabies vaccines.
Background
Rabies virus (rabbv) is the etiological agent of Rabies, which is a virulent zoonotic infectious disease characterized by infecting the central nervous system, and once the disease occurs, the lethality rate is as high as 100%. China is a high-incidence region of rabies, and the number of the people with rabies is second to that in India and is second to the world. With the increase of domestic pets, the control and prevention of rabies between people in China are still far from the greatest importance. However, the pathogenesis of rabies is still not completely clear up to now, and no specific medicine for treating rabies is available, so the most effective measure for preventing rabies is timely vaccination.
At present, rabies vaccines for human beings at home and abroad are purified protein preparations, and because of lack of corresponding adjuvants, 3-5 needles are required for the first immunization, which takes one month. As for the animal rabies vaccine, the imported inactivated vaccine has the advantages of safe use, easy storage, no pollution risk and the like. Although several inactivated vaccines with proprietary intellectual property rights are approved in China at present, due to the factors of limited production scale, high production cost and the like, the price of the inactivated vaccines is too high for the developing regions with laggard economic efficiency of Asia and Africa, so that the use of the inactivated vaccines in developing countries is limited, and most of the economically developed cities still prefer imported inactivated vaccines. Domestic veterinary inactivated vaccines need to be further reduced in cost.
Disclosure of Invention
The invention aims to overcome the defects of high cost and the like of the existing vaccine and provide the application of artesunate as an immunologic adjuvant in preparing rabies vaccine.
The invention aims to provide application of Artesunate (ART) as an active ingredient in preparation of rabies vaccine adjuvant.
ART and rabies virus vaccine strain are mixed to immunize mice, and the immune response of inactivated rabies virus in the mice is enhanced. And further identifying ART as a vaccine adjuvant and evaluating the effect of the vaccine after the ART is mixed with the vaccine, and experimental results show that the artesunate can obviously enhance the immunogenicity of the rabies vaccine and can be used as a novel rabies vaccine candidate adjuvant.
In one embodiment of the invention, the artesunate is used as the sole active ingredient.
The invention also provides a rabies vaccine adjuvant which comprises artesunate.
In one embodiment of the invention, the artesunate is used as the sole active ingredient.
The invention also provides application of the artesunate in preparation of a rabies vaccine, wherein the artesunate is used as an adjuvant.
The invention also provides a vaccine composition comprising an inactivated rabies virus strain and artesunate.
In one embodiment of the invention, the inactivated rabies virus strain is selected from at least one of CVS-11 strain, rHEP-dG strain, SAD strain, CTN-1 strain, PV strain and FluryLEP strain. Of course, not limited to the above strains, all rabies virus strains are suitable for use in the present invention.
In one embodiment of the invention, the dosage ratio of the rabies virus strain to the artesunate in the vaccine composition is 0.9 × 105～1.1×105FFU：120～130μg。
In one embodiment of the invention, the content of the rabies virus strain in the vaccine composition is 1.0X 105～1.1×105FFU：120～130μg。
In one embodiment of the invention, the amount of artesunate in the vaccine composition is 5 μ g/g animal body weight. The amount of artesunate is determined according to the weight of the animal such as mouse, and the actual concentration is 5 μ g/g mouse, for example, if each mouse is about 25g, the amount of artesunate is about 125 μ g.
In an embodiment of the present invention, the reagent further includes dimethyl sulfoxide (DMSO), and other solvents may also be used, and DMSO has a better dissolution effect when dissolving some reagents with good water solubility.
In one embodiment of the invention, in the vaccine composition artesunate (g): dimethyl sulfoxide (mL) ═ 1: (8-12), preferably 1: 10.
In one embodiment of the present invention, the buffer solution further comprises PBS buffer. The PBS buffer has the efficacy of normal saline, is more advantageous in stabilizing protein, and has the pH of 7.20-7.40.
In one embodiment of the invention, the weight ratio of artesunate to artesunate in the vaccine composition is, by volume: PBS buffer 1: (90-110), preferably 1: 100.
the invention also provides application of artesunate in preparation of rabies vaccine. The rabies vaccine is used for preventing or treating rabies of human or other animals.
Optionally, the artesunate acts as an adjuvant.
The term "adjuvant" is a pharmaceutical or immunological agent or composition that alters the efficacy of other agents, and refers in a broad sense to a broad spectrum of substances that do not provide immunity by themselves, but are capable of increasing the immunogenicity of a co-administered antigen. Adjuvants may be added to vaccines to increase the number of antibodies and long-lasting protection by increasing the immune response, thereby minimizing injected foreign substances. Adjuvants may also be used to increase the efficacy of a vaccine by helping to alter the immune response to a particular type of immune system cell, for example, by activating T cells rather than antibody-secreting B cells, depending on the purpose of the vaccine. Thus, adjuvants may advantageously modulate cytokine expression/secretion, antigen presentation, type of immune response, and the like. In the present invention, the term context refers to a compound or composition that acts as a carrier or auxiliary substance for an immunogen and/or other pharmaceutically active compound.
Wherein the above "antigen" typically refers to a substance that can be recognized by the immune system and is capable of triggering an antigen-specific immune response, e.g., by forming antibodies or antigen-specific T-cells as part of an adaptive immune response. Antigens are divided into two categories by nature: complete antigens and incomplete antigens. Complete antigen (complete antigen), abbreviated as antigen, is a substance with both immunogenicity and immunoreactivity, such as most proteins, bacteria, viruses, bacterial exotoxins, enterotoxins, and the like. An incomplete antigen, i.e. hapten (hapten), is a substance that is only immunoreactive and not immunogenic, such as most polysaccharides (e.g. capsular polysaccharide of pneumococcus), hydrolysates of capsular polysaccharides and all lipids, etc.
The invention has the following beneficial effects:
the invention firstly proposes that Artesunate (ART) which is an artemisinin derivative is taken as a rabies vaccine adjuvant.
The invention discovers that the Artesunate (ART) serving as the artemisinin derivative can obviously enhance the immunogenicity of the rabies vaccine as a rabies vaccine adjuvant through experimental research, and can serve as a novel rabies vaccine candidate adjuvant.
The research on the effect of the vaccine shows that Artesunate (ART) with lower dose can promote the immune effect of rabies vaccine and can reduce the cost for preparing the vaccine.
The vaccine effect research of the invention shows that the Artesunate (ART) which is an artemisinin derivative can generate better protective force as a rabies vaccine adjuvant.
Drawings
FIG. 1 shows the chemical formula of Artesunate (ART), an artemisinin derivative.
FIG. 2 is a graph showing the effect of ART on the body weight of KM mice, together with a PBS control group.
FIG. 3 shows a statistical graph of the level of neutralizing antibodies against rabies virus in peripheral blood after mice immunized with CVS-11 in which ART was combined with inactivation, together with a PBS control group (. about.P < 0.05).
Fig. 4 shows a statistical plot of peripheral blood anti-rabies virus neutralizing antibody levels after mice immunized with ART in combination with inactivated rHEP-dG, together with a PBS control group (. about.p < 0.05).
FIG. 5 is a graph showing the survival rate of CVS-11 challenged mice 14 days after mice immunized with ART as a vaccine adjuvant.
Detailed Description
The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
At present, the world health organization recommends the use of inactivated rabies vaccines according to the safety, and in order to develop more effective and cheaper inactivated vaccines, measures such as increasing the virus titer or searching for better adjuvants can be taken.
FIG. 1 shows the chemical formula of Artesunate (ART), an artemisinin derivative, used in the following examples, with CAS number 88495-63-0 and molecular formula: c19H28O8Molecular weight: 384.42, EINECS No.: 618-170-5.
Artesunate used in the following examples was purchased from TCI America Portland, Inc. (Tokyo chemical industry Co., Ltd.).
The artemisinin derivative Artesunate (ART) adjuvant in the following examples is prepared by mixing inactivated CVS-11 or rHEP-dG strain with ART to form rabies virus vaccine.
Specifically, the method comprises the following steps: ART was first dissolved in DMSO and then dissolved in PBS to mix with inactivated CVS-11 or rHEP-dG rabies virus when used in vivo. As per artesunate (g): the ratio of DMSO (mL) to DMSO (10: 1) is 1g, namely 1g of artesunate is dissolved in 10mL of DMSO, then the artesunate dissolved in the DMSO is dissolved in PBS buffer again, the volume ratio of artesunate to PBS buffer is 1:100, the PBS buffer is phosphate buffer, and the pH value is 7.2-7.4.
The ART is used as an adjuvant, can promote the rabies vaccine to induce to generate higher rabies virus resistant neutralizing antibody, generates better protection effect and reduces the cost of the vaccine for dogs.
Specifically, in the immunologic adjuvant, the artesunate is added into the mixture according to the dosage of 5 mu g/g mouse body weight and 1.0 multiplied by 105FFU inactivated CVS-11 or rHEP-dG, and peripheral blood was collected after intramuscular injection of immunized mice through right leg gastrocnemius and the neutralizing antibody titer against rabies virus was studied using a neutralization experiment.
In addition, the application of Artesunate (ART) as artemisinin derivative as vaccine adjuvant or in the preparation of vaccine is also within the protection scope of the present invention.
Artesunate (ART), an artemisinin derivative, was first dissolved in DMSO and then dissolved in PBS for in vivo use, mixed with inactivated CVS-11 or rHEP-dG rabies virus and used. The artesunate serving as the immune adjuvant is mixed with inactivated CVS-11 or rHEP-dG according to the dose of 5 mu g/g, serum is collected after the immune mice are injected intramuscularly through gastrocnemius muscle of the right leg, and the titer of the neutralizing antibody of the anti-rabies virus is detected by adopting a neutralization experiment. The invention also researches the biological characteristics of ART side effect, ART adjuvant vaccine immunogenicity, challenge protection and the like to evaluate the safety and effect of the ART adjuvant as a vaccine adjuvant. The result shows that ART as a novel vaccine adjuvant can not generate obvious side effect in the body of a mouse, and the ART as the adjuvant can obviously enhance the immunogenicity of the vaccine, promote the vaccine to induce the body to generate the neutralizing antibody of the anti-rabies virus with protective efficacy, and can better protect the body against the attack of the lethal rabies virus.
In terms of specific research methods, the present invention utilizes immunology related techniques to study the assessment of the efficacy of ART as a novel vaccine adjuvant in mice.
Example 1
Research on influence of novel rabies virus vaccine adjuvant ART on body weight of mice
SPF-grade Kunmin (KM) mice, 3-4 weeks old, were divided into 2 groups, namely ART group and PBS control group, each group containing 6 mice. ART was injected at a dose of 5. mu.g/g through the gastrocnemius muscle of the right leg of the mouse, and the control group was injected with an equal amount of PBS. The body weight of the mice was measured daily after dosing for a total of 15 days. The body weight ratios of all mice were compared to the body weight before each injection, and the change in body weight ratio was studied. As shown in fig. 2, the results indicate that ART does not cause weight loss in treated mice, similar to PBS treatment. More importantly, the treated mice did not show abnormal symptoms. Therefore, ART treatment at a dose of 5. mu.g/g was safe for mice.
Example 2
Immunogenicity and challenge protection research of Artesunate (ART) on rabies vaccine in KM mouse
1. Experimental methods
CVS-11 and rHEP-dG were inactivated with UV, respectively. ART was then mixed with inactivated CVS-11 or rHEP-dG, and PBS was used as an adjuvant control. SPF-level Kunming mice 6-7 weeks old were divided into 4 groups, namely ART + CVS-11 group, PBS + CVS-11 group, ART + rHEP-dG group and PBS + rHEP-dG control group, each group containing 6 mice. Experimental group Each mouse right leg gastrocnemius muscle injection 1.0X 105A mixture of FFU inactivated rabies virus and ART in a volume of 200 μ L, the amount of ART in the mixture being 5 μ g ART/g mouse body weight, artesunate (g): dimethyl sulfoxide (mL) ═ 1:10, artesunate by volume: PBS buffer 1:100, control group intramuscular injection 1.0X 105FFU inactivated rabies virus and PBS mixture. External collection on days 7 and 14 after immunizationWeekly blood, and serum anti-rabies virus neutralizing antibody levels were measured by a Fluorescent Antibody Virus Neutralization (FAVN) assay, anti-rabies standard serum purchased from the World Health Organization (WHO), the assay protocol being strictly in accordance with the World Animal Health Organization's (OIE) standard protocol.
2. Results of the experiment
As shown in fig. 3, the results indicate that immunization of mice with inactivated CVS-11 in combination with ART produced significantly higher levels of neutralizing antibodies to rabies virus than the PBS control group. As shown in figure 4, immunization of mice with inactivated hep-dG in combination with ART produced significantly higher levels of rabies virus neutralizing antibodies than the PBS control group.
The results in fig. 3 and 4 show that ART as an adjuvant for rabies vaccines is not limited to a specific inactivated rabies virus strain, but can be applied to various inactivated rabies virus strains.
3. Further, in order to verify whether the mice have protection against virulent attack after obtaining high-level immunity, live CVS-11 strain was injected intramuscularly to infect the mice at 14d after immunization, while a control group without an immune rabies vaccine was set, the morbidity and mortality of the mice were observed for 3 consecutive weeks, and the survival rate was recorded every day. As shown in fig. 5, the results showed that the mice using ART as an adjuvant had higher survival rates than the mice using PBS control.
The above results indicate that artemisinin derivative Artesunate (ART) enhances the immune response of inactivated rabies vaccine when used as an adjuvant.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (7)
1. The artesunate is used as an active ingredient for preparing rabies vaccine adjuvant.
2. Use according to claim 1, wherein artesunate is the sole active ingredient.
3. The application of the artesunate in preparing the rabies vaccine is characterized in that the artesunate is used as an adjuvant.
4. A vaccine composition, which is characterized by consisting of inactivated rabies virus strain and artesunate.
5. The vaccine composition as claimed in claim 4, wherein the inactivated rabies virus strain is selected from at least one of CVS-11 strain, rHEP-dG strain.
6. The vaccine composition of claim 4, wherein the ratio of the amount of rabies virus strain to artesunate in the vaccine composition is 0.9 x 105～1.1×105FFU：120～130μg。
7. The vaccine composition of claim 6, wherein the ratio of the amount of rabies virus strain to artesunate in the vaccine composition is 1.0 x 105～1.1×105FFU：120～130μg。
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1758905A (en) * | 2003-02-12 | 2006-04-12 | 乔治敦大学 | Use of artemisinin for treating tumors induced by oncogenic viruses and for treating viral infections |
| WO2010105096A2 (en) * | 2009-03-11 | 2010-09-16 | University Of Massachusetts | Modulation of human cytomegalovirus replication by micro-rna 132 (mir132), micro-rna 145 (mir145) and micro-rna 212 (mir212) |
| WO2014124430A1 (en) * | 2013-02-11 | 2014-08-14 | Emory University | Nucleotide and nucleoside therapeutic compositions and uses related thereto |
| CN109675027A (en) * | 2017-10-18 | 2019-04-26 | 辽宁成大生物股份有限公司 | A kind of rabies vacciness adjuvant, vaccine composition and its application |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1758905A (en) * | 2003-02-12 | 2006-04-12 | 乔治敦大学 | Use of artemisinin for treating tumors induced by oncogenic viruses and for treating viral infections |
| CN103393672A (en) * | 2003-02-12 | 2013-11-20 | 乔治敦大学 | Use of artemisinin for treating tumors induced by oncogenic viruses and for treating viral infections |
| WO2010105096A2 (en) * | 2009-03-11 | 2010-09-16 | University Of Massachusetts | Modulation of human cytomegalovirus replication by micro-rna 132 (mir132), micro-rna 145 (mir145) and micro-rna 212 (mir212) |
| WO2014124430A1 (en) * | 2013-02-11 | 2014-08-14 | Emory University | Nucleotide and nucleoside therapeutic compositions and uses related thereto |
| CN109675027A (en) * | 2017-10-18 | 2019-04-26 | 辽宁成大生物股份有限公司 | A kind of rabies vacciness adjuvant, vaccine composition and its application |
Non-Patent Citations (2)
| Title |
|---|
| LOW-LEVEL MALARIA INFECTIONS DETECTED BY A SENSITIVE POLYMERASE CHAIN REACTION ASSAY AND USE OF THIS TECHNIQUE IN THE EVALUATION OF MALARIA VACCINES IN AN ENDEMIC AREA;EGERUAN B. IMOUKHUEDE等;《AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE》;20070331;第76卷(第3期);摘要 * |
| 青蒿琥酯抗病毒作用的研究进展;李小雯等;《临床医药文献杂志》;20181029;第5卷(第87期);第190页 * |
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