CN110680911B - Japanese encephalitis vaccine soluble microneedle patch and preparation method thereof - Google Patents
Japanese encephalitis vaccine soluble microneedle patch and preparation method thereof Download PDFInfo
- Publication number
- CN110680911B CN110680911B CN201810718919.2A CN201810718919A CN110680911B CN 110680911 B CN110680911 B CN 110680911B CN 201810718919 A CN201810718919 A CN 201810718919A CN 110680911 B CN110680911 B CN 110680911B
- Authority
- CN
- China
- Prior art keywords
- vaccine
- encephalitis
- adjuvant
- needle body
- soluble microneedle
- 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.)
- Active
Links
- 229940124726 Japanese encephalitis vaccine Drugs 0.000 title claims description 27
- 238000002360 preparation method Methods 0.000 title claims description 17
- 229960005486 vaccine Drugs 0.000 claims abstract description 46
- 239000002671 adjuvant Substances 0.000 claims abstract description 42
- 206010014599 encephalitis Diseases 0.000 claims abstract description 21
- 239000011159 matrix material Substances 0.000 claims abstract description 18
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 claims abstract description 17
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 claims abstract description 17
- 239000000203 mixture Substances 0.000 claims abstract description 16
- 210000004556 brain Anatomy 0.000 claims abstract description 15
- 239000007788 liquid Substances 0.000 claims description 12
- 210000001124 body fluid Anatomy 0.000 claims description 11
- 239000010839 body fluid Substances 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 9
- 229920002674 hyaluronan Polymers 0.000 claims description 9
- 229960003160 hyaluronic acid Drugs 0.000 claims description 9
- 238000005119 centrifugation Methods 0.000 claims description 6
- 230000002441 reversible effect Effects 0.000 claims description 5
- 239000011248 coating agent Substances 0.000 claims description 4
- 238000000576 coating method Methods 0.000 claims description 4
- KPUWHANPEXNPJT-UHFFFAOYSA-N disiloxane Chemical class [SiH3]O[SiH3] KPUWHANPEXNPJT-UHFFFAOYSA-N 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 229960003239 encephalitis vaccine Drugs 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 238000007790 scraping Methods 0.000 claims description 3
- 238000000465 moulding Methods 0.000 claims description 2
- 230000001900 immune effect Effects 0.000 abstract description 20
- 238000002347 injection Methods 0.000 abstract description 5
- 239000007924 injection Substances 0.000 abstract description 5
- 230000000694 effects Effects 0.000 description 25
- 239000002131 composite material Substances 0.000 description 21
- 230000001571 immunoadjuvant effect Effects 0.000 description 17
- 239000000568 immunological adjuvant Substances 0.000 description 17
- 239000003814 drug Substances 0.000 description 16
- 229940079593 drug Drugs 0.000 description 15
- 108010042708 Acetylmuramyl-Alanyl-Isoglutamine Proteins 0.000 description 13
- BSOQXXWZTUDTEL-ZUYCGGNHSA-N muramyl dipeptide Chemical compound OC(=O)CC[C@H](C(N)=O)NC(=O)[C@H](C)NC(=O)[C@@H](C)O[C@H]1[C@H](O)[C@@H](CO)O[C@@H](O)[C@@H]1NC(C)=O BSOQXXWZTUDTEL-ZUYCGGNHSA-N 0.000 description 13
- 210000003491 skin Anatomy 0.000 description 10
- 230000001965 increasing effect Effects 0.000 description 7
- 238000011068 loading method Methods 0.000 description 7
- 239000000427 antigen Substances 0.000 description 6
- 102000036639 antigens Human genes 0.000 description 6
- 108091007433 antigens Proteins 0.000 description 6
- 230000002434 immunopotentiative effect Effects 0.000 description 6
- 239000000243 solution Substances 0.000 description 5
- 241000700159 Rattus Species 0.000 description 4
- 238000013270 controlled release Methods 0.000 description 4
- 238000012377 drug delivery Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 230000037317 transdermal delivery Effects 0.000 description 4
- 230000007423 decrease Effects 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 230000003053 immunization Effects 0.000 description 3
- 238000002649 immunization Methods 0.000 description 3
- 230000035515 penetration Effects 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 210000000434 stratum corneum Anatomy 0.000 description 3
- 238000010998 test method Methods 0.000 description 3
- 208000006400 Arbovirus Encephalitis Diseases 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 206010052369 Encephalitis lethargica Diseases 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 210000000612 antigen-presenting cell Anatomy 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 210000002615 epidermis Anatomy 0.000 description 2
- 238000010579 first pass effect Methods 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000012716 precipitator Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 201000002498 viral encephalitis Diseases 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 101100136092 Drosophila melanogaster peng gene Proteins 0.000 description 1
- 241000223932 Eimeria tenella Species 0.000 description 1
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 1
- LLPWNQMSUYAGQI-QBQUQATFSA-N Ginsenoside R1 Natural products O([C@](CC/C=C(\C)/C)(C)[C@@H]1[C@H]2[C@@H](O)C[C@H]3[C@](C)([C@]2(C)CC1)C[C@H](O[C@@H]1[C@H](O[C@@H]2[C@@H](O)[C@@H](O)[C@@H](O)CO2)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@H]1C(C)(C)[C@@H](O)CC[C@]31C)[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1 LLPWNQMSUYAGQI-QBQUQATFSA-N 0.000 description 1
- 101000746373 Homo sapiens Granulocyte-macrophage colony-stimulating factor Proteins 0.000 description 1
- 241000701076 Macacine alphaherpesvirus 1 Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 239000004411 aluminium Substances 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229940024546 aluminum hydroxide gel Drugs 0.000 description 1
- SMYKVLBUSSNXMV-UHFFFAOYSA-K aluminum;trihydroxide;hydrate Chemical compound O.[OH-].[OH-].[OH-].[Al+3] SMYKVLBUSSNXMV-UHFFFAOYSA-K 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 230000003292 diminished effect Effects 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000008098 formaldehyde solution Substances 0.000 description 1
- 229940124724 hepatitis-A vaccine Drugs 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000000091 immunopotentiator Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 238000001746 injection moulding Methods 0.000 description 1
- 210000001165 lymph node Anatomy 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000001640 nerve ending Anatomy 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- LLPWNQMSUYAGQI-OOSPGMBYSA-N notoginsenoside R1 Chemical compound O([C@@](C)(CCC=C(C)C)[C@@H]1[C@@H]2[C@@]([C@@]3(C[C@@H]([C@H]4C(C)(C)[C@@H](O)CC[C@]4(C)[C@H]3C[C@H]2O)O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O[C@H]2[C@@H]([C@@H](O)[C@H](O)CO2)O)C)(C)CC1)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O LLPWNQMSUYAGQI-OOSPGMBYSA-N 0.000 description 1
- JURZHOVRCOWZFN-UHFFFAOYSA-N notoginsenoside R1 Natural products CC(=CCCC(C)(OC1OC(CO)C(O)C(O)C1O)C2CCC3(C)C2C(O)CC4C5(C)CCC(O)C(C)(C)C5C(CC34C)OC6OC(COC7OCC(O)C(O)C7O)C(O)C(O)C6O)C JURZHOVRCOWZFN-UHFFFAOYSA-N 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000007281 self degradation Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 238000013271 transdermal drug delivery Methods 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
- A61K9/0021—Intradermal administration, e.g. through microneedle arrays, needleless injectors
-
- 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
-
- 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
- A61K2039/55522—Cytokines; Lymphokines; Interferons
-
- 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
- A61K2039/5555—Muramyl dipeptides
-
- 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
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/36011—Togaviridae
- C12N2770/36034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Virology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Dermatology (AREA)
- Molecular Biology (AREA)
- Inorganic Chemistry (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention provides a brain vaccine soluble microneedle patch which comprises a needle body and a back lining, wherein the needle body consists of a brain vaccine, a matrix material and an adjuvant, wherein the adjuvant is a mixture of MDP and GM-CSF, and the content ratio of the adjuvant to the brain vaccine is (1:1) - (1:3). The soluble micro needle of the encephalitis B vaccine prepared by the invention has good immune effect and high mechanical strength of the needle body, is hopeful to replace the encephalitis B vaccine injection and realizes painless minimally invasive administration.
Description
Technical Field
The invention relates to the technical field of microneedle administration of vaccines, and provides a soluble microneedle patch of a Japanese encephalitis vaccine and a preparation method thereof.
Background
Epidemic encephalitis B is an acute infectious disease affecting the central nervous system caused by encephalitis B virus, which often causes death of patients or leaves nervous system sequelae, and encephalitis B vaccine is an effective measure for preventing epidemic encephalitis B. The current Japanese encephalitis vaccine is generally injected and administrated through a needle, and the compliance of patients is poor, so a novel vaccine administration way is needed. In addition, protein drugs are easily decomposed by gastrointestinal tract proteases or liver first pass effects through oral administration, resulting in reduced activity, thereby affecting bioavailability. Transdermal delivery systems (TDDS) are a popular new technology for drug delivery in recent years, which can avoid the above-mentioned problems of poor patient compliance and oral digestive tract and liver first pass effects caused by needle injection, and skin is favored as the largest immune organ of the human body. However, transdermal delivery of water-soluble small molecules and macromolecular drugs has become a challenge due to the presence of skin stratum corneum disorders.
There are a variety of physical permeation promotion techniques currently available for facilitating transdermal delivery of drugs, such as microneedles, iontophoresis, electrothermic pore generation, sonophoresis, and magnetic field introduction. Other physical penetration enhancing techniques may have difficulty controlling the location and dosage of the drug administered, whereas microneedles may have their site of administration controlled by varying their length, and for soluble microneedles precise control of the dosage of the drug administered may be achieved by controlling the drug content of the cast microneedle tips, taking up the unique advantages of microneedle administration in both aspects, and thus research in the field of microneedles has increased in years. Microneedles (microscles) are array combinations with multiple micro needles that penetrate the stratum corneum of the skin into the epidermis without touching nerve endings of the dermis to achieve painless minimally invasive delivery and deliver drug effects that break through traditional transdermal drug delivery formulations. Compared with other types of microneedles, the soluble microneedles have the advantages that the drug loading is large, harmful residues cannot be caused by self-degradation after the soluble microneedles are penetrated into the skin, the sustained and controlled release can be achieved by screening different matrix material proportions, and the like, so that the soluble microneedles become one of the most studied types of the current microneedles.
There is no example of loading the Japanese encephalitis vaccine into the soluble microneedle in the current domestic and foreign patents and literature. The Japanese encephalitis vaccine is loaded into a coated microneedle for administration similarly to the Japanese encephalitis vaccine disclosed IN patent publication No. IN2008CN 0244A, and it is known that the coated microneedle and the soluble microneedle are very different IN preparation process and drug loading manner, and thus it is difficult to be referred to (Indermux S, luttge R, choonara Y E, et al Current advances IN the fabrication of microneedles for transdermal delivery [ J ]. Journal of Controlled Release,2014,185 (185): 130-138.).
An adjuvant is a nonspecific immunopotentiator that, when injected with an antigen or pre-injected into the body, enhances the body's immune response to the antigen or alters the type of immune response. The mechanism of action of immunoadjuvants can be divided into: (1) the antigen is released continuously at the site of injection (antigen-reservoir effect). (2) Up-regulate a variety of cytokines and chemokines. (3) Immune cells are recruited to the injection site. (4) Enhancing antigen uptake and presentation. (5) Activating antigen presenting cells (antigen presenting cells, APCs), promoting transport of their mature presenting antigens to draining lymph nodes. (6) Activating inflammatory corpuscles, etc. The immunopotentiation effect of different adjuvants or combinations of adjuvants on vaccines is different, SO that for different vaccines, it is necessary to screen appropriate adjuvants to achieve the corresponding immunopotentiation effect (Xu Jinjun, tao Jianping, peng Jinbiao, etc.. The effect of different adjuvants and immunization pathways on the immunoprotection effect of Eimeria tenella SO7 antigen [ J ]. Chinese Protect veterinarian journal, 2007,29 (9): 697-703; (Tao Li, duan Jinmei, shu Xiaoming, etc.. The immunopotentiation effect of notoginsenoside R1 on aluminium adjuvant hepatitis A vaccine [ J ]. J. Chinese journal of biologies, 2008,21 (3): 197-200.).
In addition to the above-mentioned advantages of soluble microneedles, there are certain limitations, such as possible insufficient mechanical strength relative to other types of microneedles. As known from the references (Park J H, allen M G, prausnitz M R. Polymer microneedles for controlled-release drug delivery [ J ]. Pharmaceutical research,2006,23 (5): 1008-1019.), the mechanical strength of the resulting soluble microneedles decreases with increasing drug ratio in the microneedle prescription. Therefore, screening for suitable microneedle drug loading ratios is of some significance in improving the mechanical strength of soluble microneedles.
In summary, how to develop a soluble microneedle with proper prescription, good immune effect and excellent mechanical strength aiming at the Japanese encephalitis vaccine is a technical problem which needs to be solved by the technicians in the field.
Disclosure of Invention
The invention aims to provide a brain vaccine soluble microneedle patch and a preparation method thereof, which solve the bottleneck existing in the prior art.
The technical scheme adopted by the invention is as follows:
the brain vaccine soluble microneedle patch comprises a needle body and a back lining, wherein the needle body consists of brain vaccine, matrix material and an adjuvant, wherein the adjuvant is the mixture of Muramyl Dipeptide (MDP) and human granulocyte macrophage colony stimulating factor (GM-CSF), and the content ratio of the adjuvant to the brain vaccine is (1:1) - (1:3).
Preferably, the matrix material is Hyaluronic Acid (HA).
Preferably, the content ratio of GM-CSF to MDP in the adjuvant is (3:1) - (1:3).
More preferably, the content ratio of GM-CSF to MDP in the adjuvant is 1:2.
preferably, the content ratio of the adjuvant to the encephalitis B vaccine is 1:2.
preferably, the matrix material accounts for 30-50% of the mass of the needle body.
Preferably, the content of each component in the needle body is as follows: the Japanese encephalitis vaccine is 40wt%, GM-CSF is 6.7wt%, MDP is 13.3wt%, and HA is 40wt%.
A preparation method of a brain vaccine soluble microneedle patch as described above, comprising the following steps:
(1) Female mold preparation of microneedles
Preparing a polydimethoxy siloxane female mould required by the brain vaccine soluble microneedle by adopting a reverse mould method;
(2) First centrifugal die-in
Mixing the components in the prescription amount, dissolving into uniform needle body fluid, coating a proper amount of needle body fluid on a female die prepared by a reverse molding method, putting into a 96-hole plate centrifuge, and centrifuging to enable the needle body fluid to enter holes of the female die; then taking out the female die, and scraping the solution outside the hole;
(3) Second centrifugation into mould
Dissolving backing into uniform backing liquid, coating a proper amount of backing liquid on the female die obtained in the step (1), putting the female die into a 96-well plate centrifuge, and centrifuging to enable the backing liquid to enter holes of the female die; and then putting the mixture into an oven, drying the mixture at 37 ℃, taking out the mixture, and demoulding the mixture to obtain the finished product.
Compared with the prior art, the brain vaccine soluble microneedle patch and the preparation method thereof provided by the invention have the following beneficial effects:
(1) the invention selects the composite immune adjuvant with specific content ratio to combine with the Japanese encephalitis vaccine, so that the immune enhancement effect of the Japanese encephalitis vaccine soluble microneedle is optimal.
(2) The invention screens proper matrix material mass ratio, so that the mechanical property of the soluble micro-needle of the Japanese encephalitis vaccine is moderate, and the administration success rate of the micro-needle is improved.
(3) The soluble microneedle prepared by the invention is a layered microneedle, so that the administration dosage of the microneedle can be controlled more easily, and the drug waste can not be caused.
In a word, the encephalitis B vaccine soluble micro-needle prepared by the invention has good immune effect and high mechanical strength, is hopeful to replace the encephalitis B vaccine injection and realizes painless minimally invasive administration.
Drawings
FIG. 1 is a schematic flow chart of a process for preparing a microneedle negative mould;
FIG. 2 is a schematic flow chart of the preparation of the Japanese encephalitis vaccine microneedle according to the present invention;
Detailed Description
In order to make the person skilled in the art better understand the solution of the present invention, the technical solution in the present embodiment will be specifically described below with reference to the accompanying drawings in the present invention. It should be noted that the following examples are only for illustrating the present invention, and not for limiting the present invention, and any modifications and changes made to the present invention within the spirit of the present invention and the scope of the appended claims fall within the scope of the present invention.
EXAMPLE 1 female preparation of brain vaccine microneedles
Referring to fig. 1, the invention adopts a reverse mould method to prepare a female mould of the Japanese encephalitis vaccine microneedle, and the steps are as follows:
the microneedle of the metal male mould is placed in a cuboid container, and the needle point is upward. The preparation method comprises the steps of (1) preparing polydimethoxyl siloxane (PDMS) and a curing agent according to a mass ratio of 10:1, and pouring the mixture into a cuboid container; placing the container in a vacuum drying oven, and respectively setting the parameter vacuum degree to be 0.07MPa and the time to be 5min to remove bubbles in the mixed solution; and then the container is put into an oven, the temperature parameter is set to be 50 ℃, the container is taken out after 5 to 8 hours, and the PDMS microneedle negative mould is obtained after demoulding.
EXAMPLE 2 preparation of Japanese encephalitis vaccine microneedle
As shown in figure 2, the invention adopts a secondary centrifugal injection molding method to prepare the Japanese encephalitis vaccine microneedle, and the steps are as follows:
1) Mixing the above components in the prescription in proportion, dissolving with appropriate amount of solvent (deionized water) to obtain uniform mixed solution (i.e. needle body fluid), loading appropriate amount of needle body fluid into centrifuge tube, centrifuging in centrifugal precipitator, centrifuging to remove bubbles in needle body fluid, and standing for use.
2) Pouring the needle body fluid obtained after the centrifugation in the step 1) on the PDMS mould prepared in the embodiment 1 of the invention, placing the PDMS mould in a table type low-speed centrifuge, and centrifuging for 5min at a rotating speed of 3000rpm so that the needle body fluid is injected into holes of the mould; after centrifugation is finished, taking out the die, scraping the solution on the surface of the die, and only keeping the solution in the holes;
3) Dissolving the composite material of the backing layer without medicine into uniform liquid (namely backing liquid) by using a solvent (deionized water), taking a proper amount of backing liquid, filling into a centrifuge tube, placing into a centrifugal precipitator, centrifuging to remove bubbles in the backing liquid, and standing for later use.
4) Pouring the backing liquid obtained after the centrifugation in the step 3) on the PDMS mould obtained in the step 2), placing the PDMS mould in a table type low-speed centrifuge, and centrifuging for 3min at a rotating speed of 3000rpm to enable the backing liquid to be injected into holes of the mould; and taking out the mould after centrifugation, putting the mould into an oven, drying at 37 ℃ for 8 hours, taking out the mould, and demoulding to obtain the finished product of the Japanese encephalitis vaccine microneedle patch.
EXAMPLE 3 Effect of the use of different immunoadjuvants on the soluble microneedle immune Effect of the Japanese encephalitis vaccine
Assuming that the ratio of the immune adjuvant in the total mass of the needle body is a certain value, only the components of the immune adjuvant are changed, and the influence of the immune adjuvants of different combinations on the immune effect of the microneedles as shown in table 1 is examined, and the preparation method of the microneedles and the female mold related to the embodiment refers to the embodiment 1 and the embodiment 2 of the invention.
TABLE 1 Effect of different immunoadjuvants on the immune Effect of soluble microneedles of Japanese encephalitis vaccine
After preparing soluble microneedles according to the methods of examples 1 and 2 of the present invention for the 11 microneedle formulations in table 1, the immune effect of the soluble microneedles was examined as follows:
the 18 soluble microneedles were penetrated into the back skin of 18 depilated rats by using a drug delivery device, and the rats were immunized again in the same manner after a period of one week after the microneedles were completely dissolved. Seven days after the secondary boost, serum was taken from the veins of the rats and the specific IgG levels of the encephalitis vaccine in the serum were quantified using an ELISA (enzyme-linked immunosorbent assay) kit.
The test results are shown in Table 2.
TABLE 2 Effect of combinations of different immunoadjuvants on the immune Effect of soluble microneedles of Japanese encephalitis vaccine
As can be seen from Table 2, the immune effect was completely different with the addition of the immune adjuvant compared with the absence of the immune adjuvant, and the immune enhancement effect of the different adjuvants on the soluble microneedles of the Japanese encephalitis vaccine was different, and the immune enhancement effect of MDP was the best when the administration was carried out with a single adjuvant, followed by MPL, GM-CSF and aluminum hydroxide gel; the use of the compound adjuvant has better immune enhancement effect than that of the single adjuvant, wherein the microneedle with the 6-size prescription corresponds to the highest serum antibody level, which shows that the immune synergy to the Japanese encephalitis vaccine is most obvious when MDP and GM-CSF adjuvants are used in combination, so that the immune adjuvant selects the mixture of MDP and GM-CSF.
Example 4 Effect of the content of the Complex immunoadjuvant on the immune Effect of the soluble microneedle of the Japanese encephalitis vaccine
On the basis of the embodiment 3, assuming that the ratio of the composite immunoadjuvant in the total mass of the needle body is a certain value and the components of the composite immunoadjuvant are fixed, only the content change of each component in the immunoadjuvant is changed, and the influence of the composite immunoadjuvant with different proportions on the microneedle immune effect is examined as shown in the table 3.
Similarly, the preparation method of the microneedle and the female mold according to the present embodiment refers to the embodiments 1 and 2 of the present invention. The immune effect of each of the soluble microneedle prescriptions in table 3 was tested and the test procedure was as described in example 3 of the present invention.
TABLE 3 Effect of various contents of composite immunoadjuvants on the immunological Effect of microneedles
The test results are shown in Table 4.
TABLE 4 Effect of different levels of composite adjuvants on the immunological effects of microneedles
As can be seen from a combination of tables 3 and 4,
1) The immunopotentiating effect is more pronounced with increasing MDP content in the composite adjuvant, but when GM-CSF: when MDP reaches 1:3 (i.e., prescription 5), the immunopotentiating effect is rather diminished.
2) When GM-CSF: mdp=1:2, the immune enhancement is optimal and therefore is the optimal prescription.
Example 5 Effect of different ratios of Complex adjuvant to vaccine on the immune Effect of microneedles
Based on example 4, assuming that the ratio of GM-CSF to MDP in the composite adjuvant is a certain value (1:2), the ratio of the matrix material is fixed, and only the ratio of the composite adjuvant to the vaccine is changed, the influence of the composite immunoadjuvant and the vaccine in different ratios on the microneedle immunity effect is examined as shown in table 5.
Similarly, the preparation method of the microneedle and the female mold according to the present embodiment refers to the embodiments 1 and 2 of the present invention. The immune effect of each of the soluble microneedle prescriptions in table 5 was tested and the test procedure was as described in example 3 of the present invention.
TABLE 5 influence of different proportions of composite immunoadjuvant and vaccine amount on the immunization effect of microneedles
| No | Composite immune adjuvant+encephalitis B vaccine | Matrix material HA (wt%) |
| 1 | Composite immunoadjuvant: encephalitis b vaccine = 1:3 | 40 |
| 2 | Composite immunoadjuvant: encephalitis b vaccine = 1:2 | 40 |
| 3 | Composite immunoadjuvant: encephalitis b vaccine = 1:1 | 40 |
The test results are shown in Table 6.
TABLE 6 influence of different proportions of composite immunoadjuvant and vaccine amount on the immunization effect of microneedles
As shown in table 6, as the ratio of the encephalitis B vaccine to the composite immune adjuvant increases from (1:1) to (2:1), the ratio of the encephalitis B vaccine increases, and the immune enhancement effect is more remarkable; when the ratio is increased to (3:1), even if the vaccine ratio continues to increase, the immunopotentiation effect is reduced with a decrease in the composite immunoadjuvant. Therefore, when the ratio of the Japanese encephalitis vaccine to the composite immune adjuvant is 2:1 (namely, prescription 2), the immune enhancement effect is optimal, so the Japanese encephalitis vaccine is taken as an optimal prescription.
EXAMPLE 6 Effect of different matrix Material ratios on microneedle Performance
As is known from the literature (Park J H, allen M G, prausnitz M R.Polymer microneedles for controlled-release drug delivery [ J ]. PharmRes,2006,23 (5): 1008-19.), the mechanical properties of microneedles are reduced with increasing drug loading in the microneedles. Thus, in addition to evaluating the immune effect, the appropriate ratio of the microneedle matrix material was determined in combination with the mechanical properties of the microneedle, and the formulation was examined as shown in table 7, assuming that the mass ratio of the composite adjuvant to the vaccine was a certain value, only the content of the matrix material was changed.
Mechanical performance test of soluble microneedles: after the microneedles were penetrated into the skin of the in vitro rats with a force of 5N using a drug applicator, the region penetrated by the skin was stained with methylene blue, and the ratio of the number of blue spots to the number of microneedles was calculated, in parallel 6 groups.
Similarly, the preparation method of the microneedle and the female mold according to the present embodiment refers to the embodiments 1 and 2 of the present invention. The immune effect of each of the soluble microneedle prescriptions in table 7 was tested, and the test procedure was as described in example 3 of the present invention.
TABLE 7 influence of different matrix Material ratios on mechanical Properties and immune Effect of microneedles
The test results are shown in Table 7, where the mechanical strength of the microneedles increases with increasing occupancy of the matrix material, the number of needles that can be administered by penetration into the skin increases, but the drug loading of the microneedles decreases with increasing occupancy. When the matrix material HA is 40% (i.e. prescription 2), the mechanical strength is moderate and the immune effect is good, so it is the optimal prescription.
In summary, the optimal prescription of the soluble microneedle of the encephalitis B vaccine is as follows: 40wt% encephalitis B vaccine, 6.7wt% GM-CSF,13.3wt%MDP,40wt%HA.
Example 7 brain vaccine soluble microneedle skin penetration experiment
After preparing soluble microneedles according to the methods of examples 1 and 2 above and the optimal prescription (40 wt% encephalitis vaccine, 6.7wt% gm-CSF,13.3wt%MDP,40wt%HA) screened in the previous examples, the skin of the isolated mice was pressed with a force of 5N using a drug applicator for 10s and then fixed in 10% formaldehyde solution, leaving histological sections. From the observation of the slicing result, the micro-needle penetrates through the stratum corneum into the epidermis layer, which shows that the soluble micro-needle prepared by the invention can effectively puncture the skin.
Claims (7)
1. The brain vaccine soluble microneedle patch comprises a needle body and a back lining, and is characterized in that the needle body consists of brain vaccine, a matrix material and an adjuvant, wherein the matrix material is hyaluronic acid; the adjuvant is a mixture of MDP and GM-CSF, and the content ratio of GM-CSF to MDP in the adjuvant is (3:1) - (1:3); the content ratio of the adjuvant to the Japanese encephalitis vaccine is (1:1) - (1:3).
2. The encephalitis B vaccine soluble microneedle patch according to claim 1, wherein the content ratio of GM-CSF to MDP in the adjuvant is 1:2.
3. the encephalitis B vaccine soluble microneedle patch of claim 1, wherein the content ratio of the adjuvant to the encephalitis B vaccine is 1:2.
4. the encephalitis B vaccine soluble microneedle patch of claim 1, wherein the matrix material accounts for 30% -50% of the mass of the needle body.
5. The encephalitis vaccine soluble microneedle patch of claim 4, wherein the matrix material comprises 40% of the mass of the needle body.
6. The encephalitis B vaccine soluble microneedle patch according to any one of claims 1 to 5, wherein the content of each component in the needle body is as follows: the Japanese encephalitis vaccine is 40wt%, GM-CSF is 6.7wt%, MDP is 13.3wt% and hyaluronic acid is 40wt%.
7. The method for preparing the encephalitis B vaccine soluble microneedle patch according to any one of claims 1 to 6, which is characterized by comprising the following steps:
(1) Female mold preparation of microneedles
Preparing a polydimethoxy siloxane female mould required by the brain vaccine soluble microneedle by adopting a reverse mould method;
(2) First centrifugal die-in
Mixing the components in the prescription amount, dissolving into uniform needle body fluid, coating a proper amount of needle body fluid on a female die prepared by a reverse molding method, putting into a 96-hole plate centrifuge, and centrifuging to enable the needle body fluid to enter holes of the female die; then taking out the female die, and scraping the solution outside the hole;
(3) Second centrifugation into mould
Dissolving backing into uniform backing liquid, coating a proper amount of backing liquid on the female die obtained in the step (1), putting the female die into a 96-well plate centrifuge, and centrifuging to enable the backing liquid to enter holes of the female die; and then putting the mixture into an oven, drying the mixture at 37 ℃, taking out the mixture, and demoulding the mixture to obtain the finished product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810718919.2A CN110680911B (en) | 2018-07-04 | 2018-07-04 | Japanese encephalitis vaccine soluble microneedle patch and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810718919.2A CN110680911B (en) | 2018-07-04 | 2018-07-04 | Japanese encephalitis vaccine soluble microneedle patch and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110680911A CN110680911A (en) | 2020-01-14 |
| CN110680911B true CN110680911B (en) | 2024-03-08 |
Family
ID=69106294
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810718919.2A Active CN110680911B (en) | 2018-07-04 | 2018-07-04 | Japanese encephalitis vaccine soluble microneedle patch and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110680911B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112516449B (en) * | 2020-12-03 | 2023-02-21 | 昆明理工大学 | Soluble microneedle array and preparation method thereof |
| CN114150022B (en) * | 2021-12-06 | 2023-08-22 | 中国科学院精密测量科学与技术创新研究院 | Biochemical molecular cell delivery method based on plant micro-nano structure and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102793917A (en) * | 2012-07-24 | 2012-11-28 | 北京中联康生物科技有限公司 | Porcine encephalitis B vaccine and preparation method thereof |
| CN107157933A (en) * | 2017-05-04 | 2017-09-15 | 同济大学 | A kind of albumen self assembly novel nano vaccine and preparation method thereof |
| CN107412943A (en) * | 2017-04-17 | 2017-12-01 | 中国人民解放军军事医学科学院微生物流行病研究所 | A kind of soluble microneedle patch and preparation method thereof |
-
2018
- 2018-07-04 CN CN201810718919.2A patent/CN110680911B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102793917A (en) * | 2012-07-24 | 2012-11-28 | 北京中联康生物科技有限公司 | Porcine encephalitis B vaccine and preparation method thereof |
| CN107412943A (en) * | 2017-04-17 | 2017-12-01 | 中国人民解放军军事医学科学院微生物流行病研究所 | A kind of soluble microneedle patch and preparation method thereof |
| CN107157933A (en) * | 2017-05-04 | 2017-09-15 | 同济大学 | A kind of albumen self assembly novel nano vaccine and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110680911A (en) | 2020-01-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR101264334B1 (en) | / solid solution perforator containing drug particle and/or drug-adsorbed particles | |
| Chen et al. | Fully embeddable chitosan microneedles as a sustained release depot for intradermal vaccination | |
| KR101747099B1 (en) | Method of Preparing Micro-Needle Using Biocompatible Polymer | |
| CN102000020B (en) | Novel micro-needle patch containing degradable polymer and preparation method thereof | |
| CN109364017B (en) | Rapid separation type soluble microneedle and preparation method thereof | |
| JP2016512754A (en) | Microarray, method of use and manufacturing for delivery of therapeutic agents | |
| US20200147209A1 (en) | Alum-containing coating formulations for microneedle vaccine patches | |
| TWI528975B (en) | Microneedle trandermal delivery device and microneedle transdermal delivery method using the same | |
| JP2016512754A5 (en) | ||
| CN105126243B (en) | Microneedle injection bullet and preparation method thereof and microneedle injection equipment | |
| WO2015122838A1 (en) | Rapidly dissolvable microneedles with drug-impregnated tips | |
| CN110680911B (en) | Japanese encephalitis vaccine soluble microneedle patch and preparation method thereof | |
| CN112516449A (en) | Soluble microneedle array and preparation method thereof | |
| CN114099414A (en) | Microneedle capable of controllably and slowly releasing medicine and preparation method thereof | |
| CN114146173B (en) | Controlled release microneedle patch and application thereof in desensitization treatment field | |
| CN108969879B (en) | Compound microneedle and microneedle patch | |
| CN205084202U (en) | Micropin injection bullet and micropin injection device | |
| CN110193137A (en) | A kind of preparation method of rabies vaccine solubility microneedle patch | |
| RU2652567C1 (en) | Microneedic applicator and method of its manufacture | |
| CN110680916B (en) | Soluble micropin of hepatitis A vaccine containing aluminum adjuvant and rofecoxib combination | |
| US20230218875A1 (en) | Particle-attached microneedle and method of manufacturing the same | |
| US20210345996A1 (en) | Minimally invasive skin biopsy method using microneedle patch | |
| CN113546166A (en) | Slow-release soluble microneedle delivery system of mite-loaded allergen microsphere | |
| CN110680798B (en) | Pneumonia polysaccharide vaccine soluble microneedle containing adjuvant MF59 and aloe extract combination | |
| CN110680908B (en) | Soluble microneedle of rapidly disintegrating type A group and C group meningococcal polysaccharide conjugate vaccine and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |