CN104274830B - A kind of nasal cavity immunity carrier based on antigen covalent bond chitosan nano - Google Patents
A kind of nasal cavity immunity carrier based on antigen covalent bond chitosan nano Download PDFInfo
- Publication number
- CN104274830B CN104274830B CN201310278891.2A CN201310278891A CN104274830B CN 104274830 B CN104274830 B CN 104274830B CN 201310278891 A CN201310278891 A CN 201310278891A CN 104274830 B CN104274830 B CN 104274830B
- Authority
- CN
- China
- Prior art keywords
- antigen
- chitosan
- nasal cavity
- covalent bond
- nanoparticle
- 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.)
- Expired - Fee Related
Links
- 102000036639 antigens Human genes 0.000 title claims abstract description 84
- 108091007433 antigens Proteins 0.000 title claims abstract description 84
- 239000000427 antigen Substances 0.000 title claims abstract description 81
- 229920001661 Chitosan Polymers 0.000 title claims abstract description 59
- 210000003928 nasal cavity Anatomy 0.000 title claims abstract description 56
- 230000036039 immunity Effects 0.000 title claims abstract description 46
- 239000002105 nanoparticle Substances 0.000 claims abstract description 54
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 16
- 229920001184 polypeptide Polymers 0.000 claims abstract description 15
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 8
- 239000000463 material Substances 0.000 claims abstract description 6
- 201000010099 disease Diseases 0.000 claims abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 5
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 4
- 238000006243 chemical reaction Methods 0.000 claims description 13
- 239000002245 particle Substances 0.000 claims description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims description 11
- -1 N-hydroxy-succinamide ester Chemical class 0.000 claims description 7
- 208000035473 Communicable disease Diseases 0.000 claims description 5
- 208000015181 infectious disease Diseases 0.000 claims description 5
- 230000000241 respiratory effect Effects 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 4
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 4
- 238000004132 cross linking Methods 0.000 claims description 4
- 125000001841 imino group Chemical group [H]N=* 0.000 claims description 4
- 206010022000 influenza Diseases 0.000 claims description 4
- 230000006196 deacetylation Effects 0.000 claims description 3
- 238000003381 deacetylation reaction Methods 0.000 claims description 3
- 238000005956 quaternization reaction Methods 0.000 claims description 3
- 238000006467 substitution reaction Methods 0.000 claims description 3
- 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 2
- 208000030507 AIDS Diseases 0.000 claims description 2
- 208000004926 Bacterial Vaginosis Diseases 0.000 claims description 2
- 206010018612 Gonorrhoea Diseases 0.000 claims description 2
- 208000001688 Herpes Genitalis Diseases 0.000 claims description 2
- 201000005505 Measles Diseases 0.000 claims description 2
- 201000009906 Meningitis Diseases 0.000 claims description 2
- 208000005647 Mumps Diseases 0.000 claims description 2
- 208000019802 Sexually transmitted disease Diseases 0.000 claims description 2
- 229920002807 Thiomer Polymers 0.000 claims description 2
- 208000007074 Trichomonas Vaginitis Diseases 0.000 claims description 2
- 208000025206 Trichomonas vaginitis urogenital infection Diseases 0.000 claims description 2
- 230000004913 activation Effects 0.000 claims description 2
- 230000010933 acylation Effects 0.000 claims description 2
- 238000005917 acylation reaction Methods 0.000 claims description 2
- 125000003368 amide group Chemical group 0.000 claims description 2
- 150000001718 carbodiimides Chemical class 0.000 claims description 2
- 230000032050 esterification Effects 0.000 claims description 2
- 238000005886 esterification reaction Methods 0.000 claims description 2
- 201000004946 genital herpes Diseases 0.000 claims description 2
- 229920000578 graft copolymer Polymers 0.000 claims description 2
- 208000002672 hepatitis B Diseases 0.000 claims description 2
- 229920002674 hyaluronan Polymers 0.000 claims description 2
- 229960003160 hyaluronic acid Drugs 0.000 claims description 2
- 230000000640 hydroxylating effect Effects 0.000 claims description 2
- 208000010805 mumps infectious disease Diseases 0.000 claims description 2
- HOGDNTQCSIKEEV-UHFFFAOYSA-N n'-hydroxybutanediamide Chemical compound NC(=O)CCC(=O)NO HOGDNTQCSIKEEV-UHFFFAOYSA-N 0.000 claims description 2
- 208000008128 pulmonary tuberculosis Diseases 0.000 claims description 2
- 201000005404 rubella Diseases 0.000 claims description 2
- 208000006379 syphilis Diseases 0.000 claims description 2
- 241000209094 Oryza Species 0.000 claims 4
- 235000007164 Oryza sativa Nutrition 0.000 claims 4
- 239000000969 carrier Substances 0.000 claims 4
- 235000013339 cereals Nutrition 0.000 claims 4
- 235000009566 rice Nutrition 0.000 claims 4
- 150000004676 glycans Chemical class 0.000 claims 1
- 210000001165 lymph node Anatomy 0.000 abstract description 11
- 210000002850 nasal mucosa Anatomy 0.000 abstract description 9
- 210000004400 mucous membrane Anatomy 0.000 abstract description 5
- 238000002360 preparation method Methods 0.000 abstract description 5
- 230000009885 systemic effect Effects 0.000 abstract description 5
- 230000004044 response Effects 0.000 abstract description 4
- 230000008685 targeting Effects 0.000 abstract description 4
- 238000005516 engineering process Methods 0.000 abstract description 3
- 230000002265 prevention Effects 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 3
- 206010057190 Respiratory tract infections Diseases 0.000 abstract description 2
- 230000000087 stabilizing effect Effects 0.000 abstract description 2
- 231100000419 toxicity Toxicity 0.000 abstract description 2
- 230000001988 toxicity Effects 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 28
- 108010058846 Ovalbumin Proteins 0.000 description 20
- 229940092253 ovalbumin Drugs 0.000 description 19
- 239000000725 suspension Substances 0.000 description 10
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 8
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 8
- 239000007995 HEPES buffer Substances 0.000 description 8
- 229960005486 vaccine Drugs 0.000 description 8
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 210000002540 macrophage Anatomy 0.000 description 6
- 241001180649 Myrcia group Species 0.000 description 5
- 210000000612 antigen-presenting cell Anatomy 0.000 description 5
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 235000019832 sodium triphosphate Nutrition 0.000 description 5
- 241001591005 Siga Species 0.000 description 4
- 230000000890 antigenic effect Effects 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 210000002751 lymph Anatomy 0.000 description 4
- 239000013641 positive control Substances 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 3
- 229910021502 aluminium hydroxide Inorganic materials 0.000 description 3
- 230000023555 blood coagulation Effects 0.000 description 3
- 229940059082 douche Drugs 0.000 description 3
- 238000010255 intramuscular injection Methods 0.000 description 3
- 239000007927 intramuscular injection Substances 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 239000000376 reactant Substances 0.000 description 3
- 210000003296 saliva Anatomy 0.000 description 3
- 239000007974 sodium acetate buffer Substances 0.000 description 3
- 230000001629 suppression Effects 0.000 description 3
- 229920002101 Chitin Polymers 0.000 description 2
- XLYOFNOQVPJJNP-ZSJDYOACSA-N Heavy water Chemical compound [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- ROOXNKNUYICQNP-UHFFFAOYSA-N ammonium persulfate Chemical compound [NH4+].[NH4+].[O-]S(=O)(=O)OOS([O-])(=O)=O ROOXNKNUYICQNP-UHFFFAOYSA-N 0.000 description 2
- 206010064097 avian influenza Diseases 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 210000004907 gland Anatomy 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 230000016379 mucosal immune response Effects 0.000 description 2
- 210000001331 nose Anatomy 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 238000001291 vacuum drying Methods 0.000 description 2
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 2
- XIVUBGOYMBSEMA-UHFFFAOYSA-N 1,4-dioxane;methylsulfinylmethane Chemical compound CS(C)=O.C1COCCO1 XIVUBGOYMBSEMA-UHFFFAOYSA-N 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 206010021703 Indifference Diseases 0.000 description 1
- 239000007987 MES buffer Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- YXYZMHGSOKYZAF-UHFFFAOYSA-M [Cl-].C(C(=C)C)(=O)OCC[N+](C(C)(C)C)(C)C Chemical class [Cl-].C(C(=C)C)(=O)OCC[N+](C(C)(C)C)(C)C YXYZMHGSOKYZAF-UHFFFAOYSA-M 0.000 description 1
- QTBSBXVTEAMEQO-GUEYOVJQSA-N acetic acid-d4 Chemical compound [2H]OC(=O)C([2H])([2H])[2H] QTBSBXVTEAMEQO-GUEYOVJQSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 229910001870 ammonium persulfate Inorganic materials 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000004700 cellular uptake Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229940099352 cholate Drugs 0.000 description 1
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
- 210000004081 cilia Anatomy 0.000 description 1
- 230000001886 ciliary effect Effects 0.000 description 1
- 238000001218 confocal laser scanning microscopy Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 238000002270 exclusion chromatography Methods 0.000 description 1
- 210000000887 face Anatomy 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 239000000568 immunological adjuvant Substances 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 208000037797 influenza A Diseases 0.000 description 1
- 229960003971 influenza vaccine Drugs 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 230000001926 lymphatic effect Effects 0.000 description 1
- 210000001365 lymphatic vessel Anatomy 0.000 description 1
- 210000003563 lymphoid tissue Anatomy 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 210000004279 orbit Anatomy 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 210000001533 respiratory mucosa Anatomy 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 210000004878 submucosal gland Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- 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
- Medicinal Preparation (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention belongs to pharmaceutical technology field, relate to a kind of nasal cavity immunity carrier based on antigen covalent bond chitosan nano and preparation method thereof；This carrier is made up of nanoparticle and antigen, and described antigen is by being covalently bonded on nanoparticle surface, and wherein, nanoparticle uses Chitosan-phospholipid complex to be prepared by material；Antigen is selected from infecting the relevant albumen of disease or tumour or polypeptide；The carrier of the present invention is remarkably improved the cervical lymph node targeting after antigen protein or polypeptide nasal-cavity administration, strengthens its mucous membrane and systemic immunity response；The preparation of described carrier is simple, the lymphonodi cervicales transhipment of process stabilizing, antigen utilization rate height, antigen difficult drop-off, beneficially raising antigen, and this carrier nasal cavity immunity is effective, without schneiderian membrane toxicity, it is adaptable to through the prevention of the disease of respiratory tract infection.
Description
Technical field
The invention belongs to pharmaceutical technology field, relate to a kind of nasal cavity immunity carrier based on antigen covalent bond chitosan nano and preparation method thereof.
Background technology
In recent years, the respiratory infectious disease caused by bacterium, fungi, virus etc. frequently occurs, as the SARS of outburst in 2003, the H5N1 bird flu broken out for 2005, the Influenza A H1N1 of 2009 are very popular and the nearest novel bird flu of H7N9 etc., the health of the mankind in serious threat；Therefore, how spreading of preventing respiratory infectious disease remains the challenge that medical personal faces.And still lack effective large area air disinfection method at present, therefore its prevention focuses on the inoculation of corresponding vaccine.
Respiratory infectious disease mainly pathogen enters or infects respiratory mucosa and causes, and schneiderian membrane is the position contacting absorbability antigenic substance at first, and therefore, nasal cavity becomes the desired site of vaccine inoculation.Nasal cavity has plurality of advantages as vaccine inoculation position: (1) inoculation is convenient, can realize nasal mucosal immune, amphi position mucosal immunity and systemic immunity simultaneously；(2) there is more Immunoactive site in nasal cavity, nose associated lymphoid tissue (NALT such as animal, be equivalent to the Waldeyer's ring of people), the wherein competent cell Han panimmunity, such as T cell, B cell, M cell, dendritic cells and macrophage etc.；(3) nasal epithelial is only made up of one layer of ciliated columnar epithelial cells, has passing through of abundant lymphatic vessel, beneficially vaccine under mucous membrane, and through lymphatic drainage to cervical lymph node, triggers immune response；(4) kind and the quantity of nasal cavity endoenzyme is far below intestines and stomach, advantageously reduces sex change or the inactivation of the big molecular antigen such as albumen, polypeptide.
But, antigenic solution direct Nasal immunization originality is relatively low, it is difficult to induction body produces long-term protection antibody, it is necessary to utilize suitable immunologic adjuvant or vaccine carrier could realize effective nasal cavity immunity；Wherein, Chitosan-phospholipid complex the nanoparticle prepared is because having good biocompatibility, can penetrate mucous epithelium, easily had been used for mucosal adjuvant by features such as antigen presenting cell (APC) identifications.
At present, described chitosan nano is to be encapsulated in inside nanoparticle by antigen as the major way of mucosal adjuvant, but this kind of method exists obvious shortcoming: (1) envelop rate is relatively low；(2) antigen is prominent releases substantially, and release is not exclusively；(3) antigen that can contact with APC and be identified by APC and offer is less so that it is immune effect is undesirable.
Therefore, currently in the urgent need to a kind of nasal cavity immunity carrier based on antigen covalent bond chitosan nano；This carrier is remarkably improved antigen utilization rate, increases the cervical lymph node targeting after antigen nasal-cavity administration, strengthens its mucous membrane and systemic immunity response.
Summary of the invention
It is an object of the invention to overcome the defect of prior art and deficiency, it is provided that a kind of new nasal cavity immunity carrier, be specifically related to a kind of nasal cavity immunity carrier based on antigen covalent bond chitosan nano and preparation method thereof；This carrier is remarkably improved antigen utilization rate, increases the cervical lymph node targeting after antigen nasal-cavity administration, strengthens its mucous membrane and systemic immunity response.
The nasal cavity immunity carrier based on antigen covalent bond chitosan nano of the present invention, it is characterized in that, it is made up of nanoparticle and antigen, described antigen is by being covalently bonded on nanoparticle surface, wherein, described nanoparticle uses Chitosan-phospholipid complex to be prepared by material, and described antigen is selected from infecting the relevant albumen of disease or tumour or polypeptide；
In the present invention, the molecular weight of described shitosan is 5000 ~ 500000, preferably 20000 ~ 200000, and deacetylation is 50% ~ 95%, preferably 70% ~ 90%；
In the present invention; described chitosan derivatives is selected from trim based on chitosan material, preferably n-trimethyl chitosan chloride and PEGization shitosans such as n-trimethyl chitosan chloride, Carboxy Chitosan, Chitosan-Thiolated Polymers, PEGization shitosan, chitosan graft polymer, hydroxylating shitosan, acylation chitosan, hyaluronic acid shitosan, esterification shitosans；
The molecular weight of described n-trimethyl chitosan chloride is 5000 ~ 600000, preferably 10000 ~ 400000；Quaternization degree is 5% ~ 60%, preferably 15% ~ 45%；
The molecular weight of described PEGization shitosan is 5000 ~ 600000, preferably 10000 ~ 300000；PEG substitution value is 3% ~ 50%, preferably 10% ~ 30%；
In the present invention, nanoparticle particle diameter prepared by described Chitosan-phospholipid complex is 50 ~ 1000 nm, preferably 50 ~ 400 nm；
In the present invention, described antigen is selected from the albumen relevant to the respiratory infectious disease such as influenza, measles, varicella, rubella, epidemic meningitis, mumps, pulmonary tuberculosis or polypeptide；
In the present invention, described antigen is selected from the albumen relevant to the sexually transmitted disease such as AIDS, hepatitis B, syphilis, gonorrhoea, trichomonas vaginitis, bacterial vaginitis, amcbiasis, genital herpes or polypeptide；
In the present invention, described antigen is selected from the albumen relevant to immunotherapy of tumors or polypeptide；
In the present invention, with the connected mode of chitosan nano, antigen includes that amido link, thioether bond, disulfide bond, ester bond etc. are any in the way of covalent bond is combined.
The present invention is covalently bound with chitin nanometer by antigen by following method:
(1) nanoparticle of shitosan or derivatives thereof is prepared with ionic cross-linking；
(2) with carbodiimide/N-hydroxy-succinamide active antigen albumen or the carboxyl of polypeptide；
(3) by the antigen hybrid reaction certain time of described nanoparticle Yu activation, by covalently bound for antigen on nanoparticle surface, prepare the covalently bound chitosan nano of antigen, obtain nasal cavity immunity carrier based on antigen covalent bond chitosan nano；
Or,
By following method, antigen is covalently bound with chitin nanometer:
(1) shitosan or derivatives thereof is reacted with 3-maleimidoproprionic acid N-hydroxy-succinamide ester, chitosan material introduces dimaleoyl imino, and then uses ionic cross-linking to prepare nanoparticle；
(2)
Antigen protein or polypeptide introduce sulfydryl；
(3)
By the antigen hybrid reaction certain time of described nanoparticle Yu sulfhydrylation, by covalently bound for antigen on nanoparticle surface, prepare the covalently bound chitosan nano of antigen, obtain nasal cavity immunity carrier based on antigen covalent bond chitosan nano.
The present invention carries out experiment in vitro, experimental result shows, described nasal cavity immunity carrier (antigen covalent bond chitosan nano), significantly enhances the macrophage intake to antigen, is 6-10 times of antigenic solution respectively, is 1.2-3 times of envelope antigen nanoparticle during 4 h.
The present invention has also carried out the nasal cavity of nasal cavity immunity carrier and has given SD rat experiment, and result shows, it is significantly concentrated on shallow table, deep cervical lymph nodes, the AUC of its lymphonodi cervicales0-4 hAnd lymph gland targeted sex index is apparently higher than the nanoparticle of envelope antigen；
After described nasal cavity immunity carrier nasal cavity immunity mouse three times, can the system of induced high levels and mucosal immune response, in blood, at IgG antibody and mucous membrane, sIgA antibody horizontal is significantly higher than the nanoparticle of envelope antigen；
Described nasal cavity immunity carrier, on nose ciliary movement, schneiderian membrane form and structure without impact, has good security.
Compared with prior art, having the prominent advantages that of this nasal cavity immunity carrier based on antigen covalent bond chitosan nano, the drawback existed when overcoming existing chitosan nano envelope antigen, is combined in antigenic molecule covalent on nanoparticle prepared by Chitosan-phospholipid complex；Its preparation method is simple, process stabilizing, and antigen utilization rate is high (up to more than 95%)；And in nasal cavity transport process, antigen difficult drop-off, be conducive to improving the lymphonodi cervicales transhipment of antigen；Described antigen is combined in nanoparticle surface, is easily identified by APC and offer after administration, strengthens immune response.Nasal cavity immunity carrier of the present invention, nasal cavity immunity is effective, without schneiderian membrane toxicity, it is adaptable to through the prevention of the disease of respiratory tract infection, have a good application prospect.
Accompanying drawing explanation
Fig. 1 is shitosan, n-trimethyl chitosan chloride and the nuclear magnetic resonance map of dimaleoyl imino-n-trimethyl chitosan chloride.
Fig. 2 shows that the form of scanning electron microscopic observation H1N1-NP and laser particle analyzer measure the particle diameter distribution situation of H1N1-NP, wherein,
A: the form of scanning electron microscopic observation H1N1-NP；
B: laser particle analyzer measures the particle diameter distribution of H1N1-NP.
Fig. 3 shows the picked-up to OVA-NP of Raw 264.7 macrophage, wherein, white arrow instruction FITC-OVA.
Fig. 4 is the activity of cervical lymph node after OVA-NP nasal-cavity administration, wherein,
A: superficial lymph is tied；
B: deep cervical lymph nodes.
Fig. 5 is mucosal sIgA antibody horizontal after OVA-NP nasal cavity immunity mouse three times, wherein,
A: nasal wash；
B: saliva；
C: vaginal douche.
Fig. 6 is the blood clotting suppression titre after H1N1-NP nasal cavity immunity mouse three times, wherein,
*:p< 0.05, there were significant differences with other each groups；
Ñ:p< 0.05, there were significant differences with HEPES solution group.
Fig. 7 is schneiderian membrane HE coloration result, wherein,
A:HEPES buffer solution (negative control)；
B:OVA-NP (5 mg/mL)；
C:OVA-NP (25 mg/mL)；
D:1% deoxysodium cholate (positive control).
Detailed description of the invention
The present invention is illustrated by following description and embodiment, is depicted below as nonrestrictive, is not limiting as scope of the presently claimed invention.
Embodiment
1
Weigh shitosan 0.1 g, be dissolved in 50 mL 1% acetic acid, the lower dropping 3-maleimidoproprionic acid hydroxysuccinimide eater solution of stirring, reacts 1 h.After reaction terminates, dialyse 24 h by reactant liquor, lyophilized, obtains dimaleoyl imino-shitosan (Mal-CS).
Weigh appropriate Mal-CS to be dissolved in sodium-acetate buffer (pH5.5), add isopyknic sodium tripolyphosphate solution, 30 mins are stirred at room temperature, prepare blank nanoparticle suspension.This suspension is mixed with sulfhydrylation ovalbumin (OVA), room temperature reaction 10
Min, obtains the covalently bound chitosan nano of OVA.
Embodiment
2
Weigh shitosan 2.5 g, be added in the acetic acid of 250 mL 1% dissolve.Add 0.10 ~ 0.14 g ammonium persulfate, fill N2Protection, adds thermal agitation, is subsequently adding 1.2 ~ 2.0 g MethacryloyloxyethylTrimethyl Trimethyl Ammonium Chlorides (TMAEMC), reacts 2 ~ 5 h.Dialyse 24 h by reactant liquor, lyophilized, obtains n-trimethyl chitosan chloride (TMC).
Weigh TMC 0.5 g, be dissolved in 50 mL sodium-acetate buffers, the lower dropping 3-maleimidoproprionic acid hydroxysuccinimide eater solution of stirring, reacts 1 h.After reaction terminates, dialyse 24 h by reactant liquor, lyophilized, obtains dimaleoyl imino-n-trimethyl chitosan chloride (Mal-TMC).
Prepared TMC, Mal-TMC are dissolved in deuterated acetic acid/heavy water,1H NMR identifies its structure, and result is as shown in Figure 1；TMC shows strong (-N at 3.2 ppm+(CH3)3) peak, it was demonstrated that TMAEMC has been connected on shitosan.Mal-TMC has obvious spike (dimaleoyl imino) to occur at 6.7 ppm, illustrates to be successfully introduced on TMC dimaleoyl imino.
Embodiment
3
Weigh appropriate Mal-TMC to be dissolved in HEPES buffer solution (pH7.0), add isopyknic sodium tripolyphosphate solution, 30 mins are stirred at room temperature, prepare blank nanoparticle suspension.This suspension is mixed with sulfhydrylation ovalbumin (OVA), room temperature reaction 10 min, obtain TMC nanoparticle covalently bound for OVA.
Embodiment
4
By appropriate mono methoxy polyethylene glycol (mPEG) and N, N-carbonyl dimidazoles (CDM) is dissolved in dioxane-dimethyl sulfoxide (2:1), 37 DEG C of stirring 18 h, it is cooled to room temperature, add absolute ether precipitated product under ice-water bath, repeatedly wash with cold dioxane-dimethyl sulfoxide-absolute ether (2:1:3), more repeatedly wash with cold absolute ether, vacuum drying, obtains mPEG-CDM.
Appropriate mPEG-CDM is dissolved in the hydrochloric acid of 0.7%, 37 DEG C of stirring 18 h, it is cooled to room temperature, with 0.5 M NaOH regulation pH7-8, ice-water bath drips acetone-absolute ether (1:2) precipitated product, cold boiling (1:10) washing precipitation, vacuum drying, obtain PEGization shitosan (mPEG-CS).
Embodiment
5
Weigh appropriate mPEG-CS to be dissolved in sodium-acetate buffer (pH5.5), add isopyknic sodium tripolyphosphate solution (1 mg/mL), 30 mins are stirred at room temperature, prepare blank nanoparticle suspension.
Appropriate ovalbumin OVA is dissolved in MES buffer solution (0.1 M; pH6.0) in; add EDC 0.4 mg and NHS 1.0 mg; 15 min are reacted under room temperature; add 1.4 mL 2 mercapto ethanol stopped reactions, this solution is mixed with blank nanoparticle suspension, under room temperature, reacts 2 h; ultrafiltration removes reaction reagent, obtains OVA covalently bound mPEG-CS nanoparticle.The joint efficiency using molecular exclusion chromatography to record OVA is 96%, and the average grain diameter of gained nanoparticle about 151nm, PDI are 0.135.
Embodiment
6
Weigh appropriate Mal-TMC to be dissolved in HEPES buffer solution (pH7.0), add isopyknic sodium tripolyphosphate solution, be stirred at room temperature and prepare blank nanoparticle suspension.This suspension is mixed with the H1N1virus antigen (H1N1) of sulfhydrylation, room temperature reaction 10 min, obtain H1N1-NP.With the form of scanning electron microscopic observation H1N1-NP, measure the particle diameter of H1N1-NP with laser particle analyzer, result as in figure 2 it is shown, H1N1-NP size is uniform, smooth rounding.Its average grain diameter is 139 nm, and PDI is 0.128, particle diameter narrow distribution.
Embodiment
7
Weigh appropriate Mal-TMC to be dissolved in HEPES buffer solution (pH7.0), add isopyknic sodium tripolyphosphate solution, be stirred at room temperature and prepare blank nanoparticle suspension.This suspension is mixed with the Influenza matrix albumen 2 film outskirt 23 peptide (M2e) being connected with sulfydryl, and in reaction system, adds the stability of Tris [2-carboxyethyl] hydrochloric acid phosphine holding polypeptide, room temperature reaction 10 min, obtain M2e-NP.With the joint efficiency of high effective liquid chromatography for measuring M2e, measure the particle diameter of nanoparticle with laser particle analyzer.M2e joint efficiency is close to 100%, and M2e-NP average grain diameter is 145
Nm, PDI are 0.119, particle diameter narrow distribution.
Embodiment
8
Use FITC mark OVA solution, the covalently bound nanoparticle of OVA (OVA-NP), the nanoparticle (NPe) of encapsulating OVA, three is hatched 1 h with macrophage Raw 264.7 respectively.The 0.04% extracellular FITC of trypan blue solution cancellation.Neutral paraformaldehyde is fixed, and DAPI dyes, and laser confocal microscope (CLSM) is observed.Result is as shown in Figure 3, it is seen that OVA-NP can be entered intracellular by macrophage picked-up, and its green fluorescence is significantly stronger than NPe.
Separately OVA solution, OVA-NP, NPe are hatched 10,30 min and 1,2,4 h, flow cytomery cell fluorescence intensity respectively with macrophage Raw 264.7.The cellular uptake amount of result OVA-NP group is the highest at each time point, is 1.99 ~ 6.71 times of OVA solution group respectively, is 1.04 ~ 1.25 times of NPe group.
Embodiment
9
SD rat is randomly divided into four groups, and wherein three groups of rat nasal cavities respectively give125The OVA solution of I mark, OVA-NP, NPe, another group (I.M. group) rat hindlimb intramuscular injection aluminium hydroxide absorption OVA vaccine, after being administered, different time puts to death rat, physiological saline cardiac perfusion, take blood, superficial lymph knot (SCLN), deep cervical lymph nodes (DCLN), weigh, intelligence is put and is exempted from γ measuring instrument and measure radiocounting, calculates activity and the lymph gland targeted sex index (LNTI) of each tissue: activity in LNTI=lymph node activity/blood.
As shown in Figure 4, after OVA-NP nasal-cavity administration, its shallow table, deep cervical lymph nodes transhipment amount are NPe groups 1.4 ~ 1.6 times to result, and its LNTI value is between 1.5 ~ 10, illustrate that it has good lymphonodi cervicales targeting.And the LNTI of I.M. group is only about 0.4, cervical lymph node transhipment amount is few.
Embodiment
10
Balb/C mouse is randomly divided into five groups, is administered once every two weeks, totally three times.Wherein four groups of mouse nasal cavities respectively give HEPES buffer solution, OVA solution, OVA-NP and NPe, and one group of intramuscular injection aluminium hydroxide absorption OVA vaccine, as positive controls.After first administration, 42 d eye sockets take blood, and collect nasal wash, saliva and vaginal douche, measure IgG, sIgA antibody horizontal in sample.
After result nasal cavity immunity three times (42d), the IgG antibody increase of OVA-NP group is the most obvious, it is significantly higher than other each group, it is OVA solution, 1192 times, 5.3 times and 2.8 times of NPe and I.M. group IgG antibody level respectively, illustrates to be connected to OVA the systemic immunity response of the mode energy induced high levels on nanoparticle surface.
After nasal-cavity administration three times, in OVA-NP group mouse nasal wash, saliva and vaginal douche, sIgA antibody horizontal the most substantially increases, and is significantly higher than other each group, and illustrating that antigen is combined on shitosan can induced high levels schneiderian membrane, amphi position mucosal immune response.Result is as shown in Figure 5.
Embodiment
11
Balb/C mouse is randomly divided into five groups, is administered once every two weeks, totally three times.Wherein four groups of mouse nasal cavities respectively give HEPES buffer solution, H1N1 solution, H1N1-NP and NPe, and one group of intramuscular injection aluminium hydroxide absorption H1N1 vaccine, as positive controls.After first administration, 42 d gather blood samples, use hemagglutination-inhibition test to evaluate whether influenza vaccines induce body to produce protection antibody.
Generally in serum, blood clotting suppression titre can regard as protected effect more than or equal to 40; from accompanying drawing 6; the blood clotting suppression titre of H1N1-NP group, NPe group and I.M. group all dramatically increases; wherein I.M. group titre value is less than 40; illustrate its still can not effectively flu-prevention virus invasion; and the titre value of H1N1-NP group and NPe group is much larger than 40, and the titre value of H1N1-NP group is significantly higher than NPe group, shows that H1N1-NP has more preferable immanoprotection action.
Embodiment
12
SD rat nasal cavity respectively gives HEPES buffer solution (negative control group), OVA-NP (5 mg/mL), OVA-NP (25
And 1% deoxycholic aicd sodium solution (positive controls) mg/mL), successive administration 3 days, last puts to death rat after being administered 24 h, separates schneiderian membrane, makees HE dyeing after paraffin section, observation by light microscope, result is as it is shown in fig. 7, the OVA-NP group schneiderian membrane of high and low concentration is complete, clear in structure, and cilium is the densest, submucosal glands, blood vessel etc. are high-visible, with negative control group indifference.As can be seen here, OVA-NP nasal cavity immunity three times on Rat Nasal Mucosa structure and cellular morphology without impact, it was demonstrated that it has good security.
Claims (14)
1. a nasal cavity immunity carrier based on antigen covalent bond chitosan nano, it is characterised in that by
Nanoparticle and antigen composition, described antigen is by being covalently bonded on nanoparticle surface；
Described nanoparticle uses Chitosan-phospholipid complex to be prepared by material；
Described antigen is selected from infecting the relevant albumen of disease or tumour or polypeptide；
Being prepared by following method, it includes step:
(1) nanoparticle of shitosan or derivatives thereof is prepared with ionic cross-linking；
(2) with carbodiimide/N-hydroxy-succinamide active antigen albumen or the carboxyl of polypeptide；
(3) by the antigen hybrid reaction certain time of described nanoparticle Yu activation, receiving covalently bound for antigen
Grain of rice surface, prepares the covalently bound chitosan nano of antigen, obtains based on antigen covalent bond chitosan nano
The nasal cavity immunity carrier of the grain of rice；
Or,
(1) shitosan or derivatives thereof is reacted with 3-maleimidoproprionic acid N-hydroxy-succinamide ester,
Chitosan material introduces dimaleoyl imino, and then uses ionic cross-linking to prepare nanoparticle；
(2) on antigen protein or polypeptide, sulfydryl is introduced；
(3) by the antigen hybrid reaction certain time of described nanoparticle Yu sulfhydrylation, receiving covalently bound for antigen
Grain of rice surface, prepares the covalently bound chitosan nano of antigen, obtains based on antigen covalent bond chitosan nano
The nasal cavity immunity carrier of the grain of rice.
2. the nasal cavity immunity carrier based on antigen covalent bond chitosan nano as described in claim 1,
It is characterized in that, the molecular weight of described shitosan is 5000～500000, and deacetylation is 50%～95%.
3. the nasal cavity immunity carrier based on antigen covalent bond chitosan nano as described in claim 1,
It is characterized in that, the molecular weight of described shitosan is 20000～200000, and deacetylation is 70%～90%.
4. the nasal cavity immunity carrier based on antigen covalent bond chitosan nano as described in claim 1, its
Be characterised by, described chitosan derivatives selected from n-trimethyl chitosan chloride, Carboxy Chitosan, Chitosan-Thiolated Polymers,
PEGization shitosan, chitosan graft polymer, hydroxylating shitosan, acylation chitosan, hyaluronic acid shell
Glycan or esterification shitosan.
5. the nasal cavity immunity carrier based on antigen covalent bond chitosan nano as described in claim 1,
It is characterized in that, described chitosan derivatives is n-trimethyl chitosan chloride and PEGization shitosan.
6. the nasal cavity immunity based on antigen covalent bond chitosan nano as described in claim 4 or 5 carries
Body, it is characterised in that the molecular weight of described n-trimethyl chitosan chloride is 5000～600000, quaternization degree is
5%～60%.
7. the nasal cavity immunity based on antigen covalent bond chitosan nano as described in claim 4 or 5 carries
Body, it is characterised in that the molecular weight of described n-trimethyl chitosan chloride is 10000～400000, quaternization degree
It is 15%～45%.
8. the nasal cavity immunity based on antigen covalent bond chitosan nano as described in claim 4 or 5 carries
Body, it is characterised in that the molecular weight of described PEGization shitosan is 5000～600000, PEG substitution value
It is 3%～50%.
9. the nasal cavity immunity based on antigen covalent bond chitosan nano as described in claim 4 or 5 carries
Body, it is characterised in that the molecular weight of described PEGization shitosan is 10000～300000, PEG substitution value
It is 10%～30%.
10. the nasal cavity immunity carrier based on antigen covalent bond chitosan nano as described in claim 1,
It is characterized in that, nanoparticle particle diameter prepared by described Chitosan-phospholipid complex is 50～1000nm.
The 11. nasal cavity immunity carriers based on antigen covalent bond chitosan nano as described in claim 1,
It is characterized in that, nanoparticle particle diameter prepared by described Chitosan-phospholipid complex is 50～400nm.
The 12. nasal cavity immunity carriers based on antigen covalent bond chitosan nano as described in claim 1,
It is characterized in that, described antigen selected from influenza, measles, varicella, rubella, epidemic meningitis, mumps,
Albumen that pulmonary tuberculosis respiratory infectious disease is relevant or polypeptide.
The 13. nasal cavity immunity carriers based on antigen covalent bond chitosan nano as described in claim 1,
It is characterized in that, described antigen selected from AIDS, hepatitis B, syphilis, gonorrhoea, trichomonas vaginitis,
Albumen that bacterial vaginitis, amcbiasis, genital herpes sexually transmitted disease are relevant or polypeptide.
The 14. nasal cavity immunity carriers based on antigen covalent bond chitosan nano as described in claim 1,
It is characterized in that, the mode that described antigen is connected with chitosan nano selected from amido link, thioether bond, disulfide bond,
Ester bond is any in the way of covalent bond combination.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310278891.2A CN104274830B (en) | 2013-07-04 | 2013-07-04 | A kind of nasal cavity immunity carrier based on antigen covalent bond chitosan nano |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310278891.2A CN104274830B (en) | 2013-07-04 | 2013-07-04 | A kind of nasal cavity immunity carrier based on antigen covalent bond chitosan nano |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104274830A CN104274830A (en) | 2015-01-14 |
| CN104274830B true CN104274830B (en) | 2016-08-17 |
Family
ID=52250490
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310278891.2A Expired - Fee Related CN104274830B (en) | 2013-07-04 | 2013-07-04 | A kind of nasal cavity immunity carrier based on antigen covalent bond chitosan nano |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104274830B (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105310986A (en) * | 2015-10-09 | 2016-02-10 | 北京万全德众医药生物技术有限公司 | Olodaterol lung targeting nanoparticle and its preparation method |
| CN105944094B (en) * | 2016-06-14 | 2019-09-13 | 中国科学院过程工程研究所 | A kind of mycobacterium tuberculosis subunit vaccine and preparation method thereof chitosan-modified based on inulin- |
| CN106267185B (en) * | 2016-09-19 | 2020-01-10 | 中国科学院过程工程研究所 | Chitosan oligosaccharide vaccine adjuvant based on chemical coupling and application thereof |
| CN106729777B (en) * | 2017-02-28 | 2020-01-14 | 华南理工大学 | Nano chitosan ultrasonic microbubble targeting prostate cancer cells and preparation and application thereof |
| CN107625961A (en) * | 2017-08-21 | 2018-01-26 | 山东大学 | The O n-trimethyl chitosan chlorides nanoparticle of sulphation Curdlan polysaccharide/6 and its application in mucosal vaccine |
| US20200306364A1 (en) * | 2017-11-30 | 2020-10-01 | Ohio State Innovation Foundation | Mucoadhesive nanoparticle entrapped influenza virus vaccine delivery system |
| CN113116854B (en) * | 2020-01-10 | 2022-07-29 | 中国药科大学 | Chitosan lipoprotein nasal administration nano-composite and preparation method and application thereof |
| WO2023168541A1 (en) * | 2022-03-09 | 2023-09-14 | Shenzhen Genius Biotech Service Co., Ltd. | Chitosan nanoparticles (cnps) and preparation method thereof |
| CN115252640B (en) * | 2022-06-23 | 2023-08-29 | 中国科学院长春应用化学研究所 | Chitosan-N-arginine nanoparticle, and preparation method and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2259914B1 (en) * | 2005-03-14 | 2007-06-16 | Advanced In Vitro Cell Technologies, S.L. | NANOPARTICULAS OF QUITOSANO AND POLYETHYLENE GLYCOL AS A SYSTEM OF ADMINISTRATION OF BIOLOGICALLY ACTIVE MOLECULES. |
| CN103083682B (en) * | 2013-01-25 | 2014-10-29 | 复旦大学 | Folic acid modified chitosan quaternary ammonium salt-taxol polymer medicine, as well as preparation method and application thereof |
-
2013
- 2013-07-04 CN CN201310278891.2A patent/CN104274830B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN104274830A (en) | 2015-01-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104274830B (en) | A kind of nasal cavity immunity carrier based on antigen covalent bond chitosan nano | |
| WO2021073659A2 (en) | Vaccine vector prepared on basis of anionic polymers and derivatives thereof | |
| Sayın et al. | TMC–MCC (N-trimethyl chitosan–mono-N-carboxymethyl chitosan) nanocomplexes for mucosal delivery of vaccines | |
| CA2894223C (en) | Nasal influenza vaccine composition | |
| Zhang et al. | Curdlan sulfate–O-linked quaternized chitosan nanoparticles: potential adjuvants to improve the immunogenicity of exogenous antigens via intranasal vaccination | |
| KR101342641B1 (en) | Composition for Adjuvant Comprising Poly-Gamma-Glutamic Acid-Chitosan Nanoparticle | |
| US10245319B2 (en) | Lymph node-targeting nanoparticles | |
| CN101845451B (en) | PEI (Polyetherimide)-chitosan triply compound gene vector with low molecular weight and preparation method and application thereof | |
| CN110237035A (en) | A kind of active targeting type Amphiphilic peptide nano-medicament carrier and its preparation and application | |
| CN104623646B (en) | A kind of double function ligand targeting dendritic cell tumor vaccine and preparation method thereof | |
| Liu et al. | Antigen-conjugated N-trimethylaminoethylmethacrylate chitosan nanoparticles induce strong immune responses after nasal administration | |
| Teng et al. | Bridging nanoplatform and vaccine delivery, a landscape of strategy to enhance nasal immunity | |
| Marin et al. | Next generation polyphosphazene immunoadjuvant: Synthesis, self-assembly and in vivo potency with human papillomavirus VLPs-based vaccine | |
| CN108785666A (en) | A kind of DC targeted nanometer vaccines and preparation method thereof applied to liver cancer immunity treatment | |
| CN102327228A (en) | Preparation method of chitosan newcastle disease vaccine nanoparticles | |
| CN103627005A (en) | Polyethylene glycol modified polyethyleneimine and use of polyethylene glycol modified polyethyleneimine as antigen protein vector | |
| ES2878071T3 (en) | New immunobiological products | |
| CN107200788B (en) | Quaternary phosphonium chitosan and application thereof as vaccine immunologic adjuvant | |
| CN105919937B (en) | A kind of nano suspension and preparation method thereof being immunized for oral protein | |
| KR102631084B1 (en) | Complex for enhancing immune response | |
| RU2779622C9 (en) | Complex for enhancing immune response | |
| EP4316511A1 (en) | Nanogel-coated vaccine | |
| Janovska et al. | CHITOSAN NANOPARTICLES AS DELIVERY SYSTEM FOR NASAL IMMUNISATION. | |
| CN116726161A (en) | Albumin delivery system loaded with antigen polypeptide and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160817 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |