CN103908668A - 蝇蛆抗菌肽佐剂及其制备方法和含该佐剂的疫苗制剂及用途 - Google Patents
蝇蛆抗菌肽佐剂及其制备方法和含该佐剂的疫苗制剂及用途 Download PDFInfo
- Publication number
- CN103908668A CN103908668A CN201410133703.1A CN201410133703A CN103908668A CN 103908668 A CN103908668 A CN 103908668A CN 201410133703 A CN201410133703 A CN 201410133703A CN 103908668 A CN103908668 A CN 103908668A
- Authority
- CN
- China
- Prior art keywords
- adjuvant
- antibacterial peptide
- fly larvae
- preparation
- vaccine
- 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.)
- Granted
Links
- 239000003910 polypeptide antibiotic agent Substances 0.000 title claims abstract description 69
- 239000002671 adjuvant Substances 0.000 title claims abstract description 66
- 238000002360 preparation method Methods 0.000 title claims abstract description 35
- 229960005486 vaccine Drugs 0.000 title abstract description 47
- 239000000203 mixture Substances 0.000 title abstract description 12
- 238000009472 formulation Methods 0.000 title abstract 3
- 238000000034 method Methods 0.000 claims abstract description 6
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 5
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 5
- 229920001184 polypeptide Polymers 0.000 claims abstract description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 22
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 18
- 239000000284 extract Substances 0.000 claims description 12
- 239000011347 resin Substances 0.000 claims description 7
- 229920005989 resin Polymers 0.000 claims description 7
- 238000004108 freeze drying Methods 0.000 claims description 6
- 239000003463 adsorbent Substances 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 4
- 238000010828 elution Methods 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 238000000746 purification Methods 0.000 claims description 3
- 230000000274 adsorptive effect Effects 0.000 claims description 2
- 239000002398 materia medica Substances 0.000 claims 1
- 241000699670 Mus sp. Species 0.000 abstract description 13
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 abstract description 12
- 229910052782 aluminium Inorganic materials 0.000 abstract description 12
- 239000000568 immunological adjuvant Substances 0.000 abstract description 10
- 230000001900 immune effect Effects 0.000 abstract description 4
- 230000007969 cellular immunity Effects 0.000 abstract 1
- 230000028996 humoral immune response Effects 0.000 abstract 1
- 230000036046 immunoreaction Effects 0.000 abstract 1
- 108010058846 Ovalbumin Proteins 0.000 description 23
- 229940092253 ovalbumin Drugs 0.000 description 23
- 241000699666 Mus <mouse, genus> Species 0.000 description 21
- 210000004027 cell Anatomy 0.000 description 20
- UZQJVUCHXGYFLQ-AYDHOLPZSA-N [(2s,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-4-[(2r,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-3,5-dihydroxy-6-(hydroxymethyl)-4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-3,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3,5-dihydroxy-6-(hy Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O)O[C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O)O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2[C@@]1(C=O)C)C)(C)CC(O)[C@]1(CCC(CC14)(C)C)C(=O)O[C@H]1[C@@H]([C@@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O[C@H]4[C@@H]([C@@H](O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O)[C@H](O)[C@@H](CO)O4)O)[C@H](O)[C@@H](CO)O3)O)[C@H](O)[C@@H](CO)O2)O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UZQJVUCHXGYFLQ-AYDHOLPZSA-N 0.000 description 18
- BLWINLJDTOJSRU-UHFFFAOYSA-N 2-methoxy-2,4-diphenylfuran-3-one Chemical compound O=C1C(OC)(C=2C=CC=CC=2)OC=C1C1=CC=CC=C1 BLWINLJDTOJSRU-UHFFFAOYSA-N 0.000 description 16
- 241000257159 Musca domestica Species 0.000 description 16
- 239000004411 aluminium Substances 0.000 description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 10
- 230000036039 immunity Effects 0.000 description 10
- 239000000427 antigen Substances 0.000 description 8
- 102000036639 antigens Human genes 0.000 description 8
- 108091007433 antigens Proteins 0.000 description 8
- 239000003292 glue Substances 0.000 description 8
- 238000011160 research Methods 0.000 description 8
- 239000003085 diluting agent Substances 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 239000012646 vaccine adjuvant Substances 0.000 description 7
- 229940124931 vaccine adjuvant Drugs 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 230000028327 secretion Effects 0.000 description 6
- 210000002966 serum Anatomy 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 230000000240 adjuvant effect Effects 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 239000002158 endotoxin Substances 0.000 description 5
- 230000006698 induction Effects 0.000 description 5
- 239000007924 injection Substances 0.000 description 5
- 238000002347 injection Methods 0.000 description 5
- 229920006008 lipopolysaccharide Polymers 0.000 description 5
- 108010062580 Concanavalin A Proteins 0.000 description 4
- 108010002350 Interleukin-2 Proteins 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000003053 immunization Effects 0.000 description 4
- 238000002649 immunization Methods 0.000 description 4
- 230000001571 immunoadjuvant effect Effects 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 210000004988 splenocyte Anatomy 0.000 description 4
- 230000000638 stimulation Effects 0.000 description 4
- 201000001320 Atherosclerosis Diseases 0.000 description 3
- 108090000695 Cytokines Proteins 0.000 description 3
- 102000004127 Cytokines Human genes 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 239000000839 emulsion Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000028993 immune response Effects 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 229940124872 Hepatitis B virus vaccine Drugs 0.000 description 2
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 2
- 229910021502 aluminium hydroxide Inorganic materials 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 230000000840 anti-viral effect Effects 0.000 description 2
- 230000000890 antigenic effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000010241 blood sampling Methods 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 230000006837 decompression Effects 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000002519 immonomodulatory effect Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000008176 lyophilized powder Substances 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 238000010254 subcutaneous injection Methods 0.000 description 2
- 239000007929 subcutaneous injection Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- LAOOXBLMIJHMFO-UHFFFAOYSA-N 1-[2-(diethylamino)ethylamino]-4-methylthioxanthen-9-one;hydron;chloride Chemical compound Cl.S1C2=CC=CC=C2C(=O)C2=C1C(C)=CC=C2NCCN(CC)CC LAOOXBLMIJHMFO-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 101710186708 Agglutinin Proteins 0.000 description 1
- 206010002198 Anaphylactic reaction Diseases 0.000 description 1
- 101710120040 Antifungal peptide Proteins 0.000 description 1
- 108050004290 Cecropin Proteins 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 238000011238 DNA vaccination Methods 0.000 description 1
- 241000255925 Diptera Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 206010018691 Granuloma Diseases 0.000 description 1
- 241000711549 Hepacivirus C Species 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 101000669447 Homo sapiens Toll-like receptor 4 Proteins 0.000 description 1
- 101710146024 Horcolin Proteins 0.000 description 1
- 208000015817 Infant Nutrition disease Diseases 0.000 description 1
- 101710189395 Lectin Proteins 0.000 description 1
- 206010067125 Liver injury Diseases 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 101710179758 Mannose-specific lectin Proteins 0.000 description 1
- 101710150763 Mannose-specific lectin 1 Proteins 0.000 description 1
- 101710150745 Mannose-specific lectin 2 Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001092142 Molina Species 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 241000257229 Musca <genus> Species 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 201000005702 Pertussis Diseases 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 241001454523 Quillaja saponaria Species 0.000 description 1
- 235000009001 Quillaja saponaria Nutrition 0.000 description 1
- 206010037888 Rash pustular Diseases 0.000 description 1
- 241000219287 Saponaria Species 0.000 description 1
- 241000700584 Simplexvirus Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 102100039360 Toll-like receptor 4 Human genes 0.000 description 1
- 208000037386 Typhoid Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000000910 agglutinin Substances 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229940037003 alum Drugs 0.000 description 1
- AZDRQVAHHNSJOQ-UHFFFAOYSA-N alumane Chemical class [AlH3] AZDRQVAHHNSJOQ-UHFFFAOYSA-N 0.000 description 1
- 230000036783 anaphylactic response Effects 0.000 description 1
- 208000003455 anaphylaxis Diseases 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- CJZGTCYPCWQAJB-UHFFFAOYSA-L calcium stearate Chemical compound [Ca+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O CJZGTCYPCWQAJB-UHFFFAOYSA-L 0.000 description 1
- 235000013539 calcium stearate Nutrition 0.000 description 1
- 239000008116 calcium stearate Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 229940028617 conventional vaccine Drugs 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 208000001848 dysentery Diseases 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 230000007760 free radical scavenging Effects 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 210000000087 hemolymph Anatomy 0.000 description 1
- 231100000234 hepatic damage Toxicity 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 1
- 230000002443 hepatoprotective effect Effects 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000000091 immunopotentiator Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000007688 immunotoxicity Effects 0.000 description 1
- 231100000386 immunotoxicity Toxicity 0.000 description 1
- 230000005918 in vitro anti-tumor Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000008818 liver damage Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 201000004792 malaria Diseases 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005360 mashing Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000002088 nanocapsule Substances 0.000 description 1
- 239000002077 nanosphere Substances 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 229940023041 peptide vaccine Drugs 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000007112 pro inflammatory response Effects 0.000 description 1
- 208000029561 pustule Diseases 0.000 description 1
- 150000003856 quaternary ammonium compounds Chemical class 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 235000017709 saponins Nutrition 0.000 description 1
- 230000001843 schistosomicidal effect Effects 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000008279 sol Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007962 solid dispersion Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 201000008297 typhoid fever Diseases 0.000 description 1
- 241000712461 unidentified influenza virus Species 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
- 230000005924 vaccine-induced immune response Effects 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 238000004804 winding Methods 0.000 description 1
Abstract
本发明涉及具有免疫佐剂作用的蝇蛆抗菌肽及其制备方法和含该抗菌肽佐剂的疫苗制剂,以及它们在制备疫苗制剂中的应用。该抗菌肽是从蝇蛆中提取分离的多肽组分,可促进免疫小鼠的细胞免疫和体液免疫应答,诱生机体同时产生平衡的Th1型和Th2型免疫反应,显示出比现有技术中已知的铝胶佐剂更佳的佐剂特性,可以作为多种疫苗的免疫佐剂,发挥理想的免疫效果。以该抗菌肽为佐剂的疫苗制备工艺简单、方法简便、质量容易控制、使用方便、可冰冻保存,安全性好。
Description
技术领域
本发明涉及从蝇蛆提取分离的具有免疫佐剂作用的蝇蛆抗菌肽,及其制备方法和含该抗菌肽佐剂的疫苗制剂,以及它们在制备疫苗制剂中的应用。
背景技术
疫苗接种是预防和控制传染性疾病最经济、有效的手段。DNA疫苗、重组疫苗和合成肽疫苗等新型疫苗是近年来发展迅速的一类生物制剂,比常规疫苗具有抗原纯度高、特异性强、安全性好等优势。然而,这些新型疫苗的免疫原性弱,免疫效力低下[Oyston P, Robinso K. The current challenges for vaccine development. J Med Microbiol 2012; 61(7): 889–894]。同此,当前疫苗研究正面临一些复杂病原如疟疾、结核、人免疫缺陷病毒等的重要挑战[Leroux–Roels G. Unmet needs in modern vaccinology: Adjuvants to improve the immune response. Vaccine 2010; 28(Supp. 3): C25–36]。佐剂是传统疫苗和当前新型疫苗必不可少的组成成分,不仅能影响机体对疫苗免疫应答的强度,而且能针对特定病原体诱导最有效的免疫应答类型[Mbow ML, De Gregorio E, Valiante NM, et al. New adjuvants for human vaccines. Current Opinion in Immunology 2010; 22(3): 411–416]。目前研究的免疫佐剂种类很多,如油佐剂、弗氏佐剂、微生物及其代谢产物、核酸及其类似物、细胞因子、脂质体等[Dey AK, Srivastava IK. Novel adjuvants and delivery systems for enhancing immune responses induced by immunogens. Expert Rev Vaccines 2011; 10: 227–51],但由于存在不同程度的毒副作用或安全隐患等一些不可避免的缺陷而难以实际应用[Mbow ML, De Gregorio E, Valiante NM, Rappuoli R. New adjuvants for human vaccines. Curr Opin Immunol 2010, 22(3): 411–6; Batista–Duharte A, Lindblad EB, Oviedo–Orta E. Progress in understanding adjuvant immunotoxicity mechanisms. Toxicol Lett 2011; 203(2): 97–105]。铝胶佐剂自1920s使用以来,直至最近由铝盐和TLR4激动剂3–O–去酰基–4’–单磷酰脂质A组成的AS04被批准之前一直是唯一通过FDA批准的人用疫苗佐剂[Marrack P, McKee AS, Munks MW. Towards an understanding of the adjuvant action of aluminium. Nat Rev Immunol 2009; 9: 287–293]。然而,铝胶佐剂存在一些弊端:① 主要通过诱导Th2型免疫应答产生抗体起保护作用,不能诱生Th1型免疫应答和细胞介导的免疫反应,只适用于以抗体为主要保护性免疫的疫苗(Exley C, Siesjo P, Eriksson H. The immunobiology of aluminium adjuvants: how do they really work? Trends Immunol 2010; 31: 103–109);② 对人免疫缺陷病毒、丙型肝炎病毒、单纯疱疹病毒、流感病毒以及血吸虫病、百日咳和伤寒等多种抗原无佐剂作用[O’Hagan DT, De Gregorio E. The path to a successful vaccine adjuvant- ‘the long and winding road’ . Drug Discov Today 2009; 14: 541–551];③ 可促进IgE抗体产生,容易诱导机体产生过敏反应;④ 在局部形成肉芽肿,甚至发生局部无菌性脓肿;⑤ 因其理化性质特点,含铝胶的疫苗怕冻,冻后铝胶容易变性;⑥ 对人、畜神经系统可能有影响。因此,铝胶佐剂已远远不能满足新型疫苗发展的要求[Kool M, Fierens K, Lambrecht BN. Alum adjuvant: some of the tricks of the oldest adjuvant. J Med Microbiol 2012; 61(7): 927–934]。寻找安全、有效、新型的免疫佐剂是当前疫苗研究领域的一个热点[Schijns VEJC, Lavelle EC. Trends in vaccine adjuvants. Expert Rev Vaccines 2011; 10(4): 539–550]。免疫佐剂研究已被列为疫苗研究的优先领域[Harandi AM, Medaglini D, Shattock RJ. Vaccine adjuvants: A priority for vaccine research. Vaccine 2010; 28(12): 2363–2366; Harand AM?, Brewe J, Schijn V. Conference Scene: Recent advancements in immunopotentiators for modern vaccines. Immunotherapy 2011; 3(11): 1297–301]。
天然药物用于防治疾病具有悠久的历史。当今,FDA批准的药物约30%来源于天然药物。新的具有免疫活性的天然产物的发现日益成为研究重点,尤其在寻找新一代疫苗佐剂方面[Licciardi PV, Underwood JR. Plant–derived medicines: A novel class of immunological adjuvants. Int Immunopharmacol 2011; 11(3): 390–398]。
蝇蛆为昆虫纲双翅目环裂亚目家蝇科家蝇Musca domestica L.的幼虫。蝇蛆在我国食用和药用历史悠久,主治臁烂、唇疔、毒痢作呕、小儿疳积诸症。蝇蛆的主要化学成分包括蛋白质、抗菌肽、不饱和脂肪酸、多糖、溶菌酶、凝集素、维生素和矿物质等[Feng X, Cheng G, Chen SY, Yang H, Huang W. Evaluation of the burn healing properties of oil extraction from housefly larva in mice. J Ethnopharmacol 2010; 130(3): 586–92]。蝇蛆抗菌肽具有抗氧化[Ai H, Wang F, Lei C. Antioxidant activities of protein–enriched fraction from the larvae of housefly, Musca domestica. Nat Prod Res 2008; 22(6): 507–15]、抗肿瘤[Hou L, Shi Y, Zhai P, Le G. Antibacterial activity and in vitro anti–tumor activity of the extract of the larvae of the housefly (Musca domestica). J Ethnopharmacol 2007; 111(2): 227–31; Jin XB, Mei HF, Li XB, Ma Y, Zeng AH, Wang Y, et al. Apoptosis–inducing activity of the antimicrobial peptide cecropin of Musca domestica in human hepatocellular carcinoma cell line BEL–7402 and the possible mechanism. Acta Biochim Biophys Sin 2010; 42(4): 259–65; Zhu L, Wang P, Qin QL, Zhang H, Wu YJ. Protective effect of polypeptides from larva of housefly (Musca domestica) on hydrogen peroxide–induced oxidative damage in HepG2 cells. Food Chem Toxicol 2013; 60: 385–90]、抗真菌[Fu P, Wu J, Guo G. Purification and molecular identification of an antifungal peptide from the hemolymph of Musca domestica (housefly). Cell Mol Immunol 2009; 6(4): 245–51.]、抗炎[Chu FJ, Jin XB, Xu YY, Ma Y, Li XB, Lu XM, et al. Inflammatory regulation effect and action mechanism of anti–inflammatory effective parts of housefly (Musca domestica) larvae on atherosclerosis. Evid–based Complement Altern Med 2013; 2013 :Article ID 340267, 10 pages]、抗动脉粥样硬化[Chu FJ, Jin XB, Zhu JY. Housefly maggots (Musca domestica) protein–enriched fraction/extracts (PE) inhibit lipopolysaccharide–induced atherosclerosis pro–inflammatory responses. J Atheroscler Thromb 2011; 18(4): 282–90]、保肝[Wang FR, Ai H, Chen XM, Lei CL. Hepatoprotective effect of a protein–fraction from the maggots (Musca domestica) against CCl4–induced hepatic damage. Biotechnol Letter 2007; 29: 853–8]、抗病毒以及免疫调节[Ai H, Wang F, Zhang N, Zhang L, Lei C. Antiviral, immunomodulatory, and free radical scavenging activities of a protein–enriched fraction from the larvae of the housefly, Musca domestica. J Insect Sci 2013; 13:112.]等活性。然而,迄今蝇蛆抗菌肽作为疫苗佐剂的研究尚未见报道。
发明内容
本发明的目的是提供一类用于疫苗、具有免疫佐剂作用的蝇蛆抗菌肽佐剂。
本发明的另一个目的是提供从蝇蛆中提取分离蝇蛆抗菌肽佐剂的方法。
本发明的进一步目的是提供一种包含所述蝇蛆抗菌肽佐剂的疫苗制剂。
本发明的还有一个目的是提供所述蝇蛆抗菌肽佐剂在制备疫苗制剂中的应用。
本发明的蝇蛆抗菌肽佐剂是从蝇蛆中提取、分离的蝇蛆多肽组分。蝇蛆抗菌肽中蛋白质的质量百分含量为56.24%±3.9%,分子量约10 KDa。
本发明提供的蝇蛆抗菌肽是从蝇蛆中提取分离的,制备方法包括以下步骤:
a. 将蝇蛆以质量浓度3%醋酸提取;
b. 醋酸提取液100℃加热1 h,4℃静置,滤过;
c. 滤液浓缩,冷冻干燥,得蝇蛆提取物;
d. 蝇蛆提取物上大孔吸附树脂柱进行色谱纯化,以水和55%乙醇依次洗脱,收集55%乙醇洗脱液,浓缩,冷冻干燥,得蝇蛆抗菌肽。
上述步骤d中所说的大孔吸附树脂为D101大孔吸附树脂。
本发明的含蝇蛆抗菌肽佐剂的疫苗制剂,包含有免疫有效量的蝇蛆抗菌肽佐剂以及含有一种或多种药物学上可接受的载体。
本发明的蝇蛆抗菌肽佐剂可在制备疫苗制剂中应用。
上文所述药学上可接受的载体是指药学领域的药物载体。例如:稀释剂、赋形剂如水、生理盐水、葡萄糖、甘露醇、甘油、乙醇及其混合物等;填充剂如淀粉、蔗糖等;粘合剂如纤维素衍生物、海藻酸盐、明胶和聚乙烯吡咯烷酮;湿润剂如甘油;崩解剂如碳酸钙和碳酸氢钠;吸收促进剂如季铵化合物;表面活性剂如吐温–80;润滑剂如滑石、硬脂酸钙、硬脂酸镁和聚乙二醇等。另外还可以在组合物中加入其它辅料如香味剂、甜味剂等。
本发明的疫苗制剂可以是溶液、混悬液或乳浊液形式,可通过口服、鼻吸入、直肠、肠胃外或经皮给药的方式施用于需要接种的对象。本发明的疫苗制剂用于口服时,可将其制成常规的固体制剂如片剂、粉剂、颗粒剂、胶囊剂、丸剂、缓释微丸、固体分散体、包含物等,制成的液体制剂如混悬剂、乳剂、溶胶剂、糖浆剂、合剂、溶液剂等;用于肠胃外给药时,可将其制成注射用的溶液、水或油性混悬剂、乳剂、冻干粉、脂质体、微囊、微球、纳米囊、纳米球等。优先的形式是注射用冻干粉及溶液。
本发明的疫苗制剂可按照药学和疫苗领域的任何常规生产方法制备。
疫苗制剂的典型单剂量可以根据制剂的形式而变化,并根据抗原种类、所需的抗体水平、接种对象的特异性及所需的免疫程序等各种因素进行确定。本发明的疫苗制剂中蝇蛆抗菌肽佐剂的量可为0.0 1 μg~1 g /单剂量,优选为0.1~100 μg/单剂量。可以一次或多次施用。
本发明的优点
本发明具有免疫佐剂作用的蝇蛆抗菌肽可促进免疫小鼠的细胞免疫和体液免疫应答,诱生机体同时产生平衡的Th1型和Th2型免疫反应,显示出比现有技术中已知的铝胶佐剂更佳的佐剂特性,可以作为多种疫苗的免疫佐剂,发挥理想的免疫效果。以该抗菌肽为佐剂的疫苗制备工艺简单、方法简便、质量容易控制、使用方便、可冰冻保存,安全性好。。所以,本发明的蝇蛆抗菌肽有望开发成疫苗佐剂。
附图说明
图1为蝇蛆抗菌肽(MDPF)对卵清蛋白(OVA)受免小鼠血清中特异性IgG、IgG1、IgG2a和IgG2b抗体效价的影响。Quil A:佐剂对照,系从南美皂树(Quillaja saponaria Molina)树皮中分离制备的佐剂活性总皂苷。a P<0.05、b P<0.01和c P<0.001 vs OVA对照组。
图2为蝇蛆抗菌肽(MDPF)对Con A刺激的卵清蛋白(OVA)受免小鼠脾细胞增殖反应的影响。Quil A:佐剂对照。a P<0.05和b P<0.01 vs OVA对照组。
图3为蝇蛆抗菌肽(MDPF)对LPS刺激的卵清蛋白(OVA)受免小鼠脾细胞增殖反应的影响。Quil A:佐剂对照。a P<0.和、b P<0.01 vs OVA对照组组。
图4为蝇蛆抗菌肽(MDPF)对卵清蛋白(OVA)受免小鼠脾细胞分泌IL-2能力的影响。Quil A:佐剂对照。c P<0.001 vs OVA对照组。
图5为蝇蛆抗菌肽(MDPF)对卵清蛋白(OVA)受免小鼠脾细胞分泌INF-γ能力的影响。Quil A:佐剂对照。a P<0.05、b P<0.01和c P<0.001 vs OVA对照组。
图6为蝇蛆抗菌肽(MDPF)对卵清蛋白(OVA)受免小鼠脾细胞分泌IL-10能力的影响。Quil A:佐剂对照。c P<0.001 vs OVA对照组。
图7为蝇蛆抗菌肽(MDPF)对禽流感–新城疫重组二联活疫苗(rL-H5)受免小鼠血清中特异性IgG、IgG1、IgG2a和IgG2b抗体效价的影响。Quil A:佐剂对照。a P<0.05、b P<0.01和c P<0.001 vs rL-H5对照组。
图8为蝇蛆抗菌肽(MDPF)对Con A刺激的禽流感–新城疫重组二联活疫苗(rL-H5)受免小鼠脾细胞增殖反应的影响。Quil A:佐剂对照。a P<0.05、b P<0.01和c P<0.001 vs rL-H5对照组。
图9为蝇蛆抗菌肽(MDPF)对LPS刺激的禽流感–新城疫重组二联活疫苗(rL-H5)受免小鼠脾细胞增殖反应的影响。Quil A:佐剂对照。a P<0.05、b P<0.01和c P<0.001 vs rL-H5对照组。
图10为蝇蛆抗菌肽(MDPF)对H5抗原刺激的禽流感–新城疫重组二联活疫苗(rL-H5)受免小鼠脾细胞增殖反应的影响。Quil A:佐剂对照。a P<0.05、b P<0.01和c P<0.001 vs rL-H5对照组。
图11为蝇蛆抗菌肽(MDPF)对禽流感–新城疫重组二联活疫苗(rL-H5)受免小鼠自然杀伤(NK)细胞活性的影响。Quil A:佐剂对照。a P<0.05、b P<0.01和c P<0.001 vs rL-H5对照组。
图12为蝇蛆抗菌肽(MDPF)对禽流感–新城疫重组二联活疫苗(rL-H5)受免小鼠脾细胞分泌IL-2能力的影响。Quil A:佐剂对照。b P<0.01和c P<0.001 vs rL-H5对照组。
图13为蝇蛆抗菌肽(MDPF)对禽流感–新城疫重组二联活疫苗(rL-H5)受免小鼠脾细胞分泌INF-γ能力的影响。Quil A:佐剂对照。c P<0.001 vs rL-H5对照组。
图14为蝇蛆抗菌肽(MDPF)对禽流感–新城疫重组二联活疫苗(rL–H5)受免小鼠脾细胞分泌IL-10能力的影响。Quil A:佐剂对照。c P<0.001 vs rL-H5对照组。
具体实施方式
以下通过实例进一步说明本发明,而非限制其范围。
以下百分比浓度均指质量百分比浓度。
实施例1:蝇蛆抗菌肽制备
取蝇蛆1 kg,以3%醋酸水溶液于高速组织捣碎机中匀浆5 min,置4℃冰箱中放置过夜,离心,收集上清液;沉淀再以3%醋酸水溶液同法处理。合并两次上清液, 100 ℃加热1 h,4℃冰箱中静置,减压抽滤。滤液减压浓缩,冷冻干燥、得到蝇蛆提取物。蝇蛆提取物上D101大孔吸附树脂色谱柱,用水和55%乙醇依次进行洗脱,收集55%乙醇洗脱液,减压回收乙醇、浓缩,冷冻干燥,得蝇蛆抗菌肽。蝇蛆抗菌肽为浅黄白色粉末;茚三酮反应呈阳性;蛋白质质量百分含量为56.24%±3.9%;分子量约10 KDa。
实施例2 含蝇蛆抗菌肽佐剂的乙肝疫苗制剂
称取蝇蛆抗菌肽500 μg和重组乙型肝炎病毒表面抗原(rHBsAg)100μg,以生理盐水10 ml溶解,以0.22 μm微孔滤膜滤过,无菌分装,每支1 ml。每ml含抗菌肽50 μg和rHBsAg 10 μg。
实施例3 含铝胶和蝇蛆抗菌肽佐剂的乙肝疫苗制剂
称取蝇蛆抗菌肽适量,以生理盐水溶解,制成浓度为100μg/ml的溶液,以0.22μm微孔滤膜滤过。将该抗菌肽溶液与等体积的含铝佐剂乙型肝炎病毒表面抗原疫苗原液(每ml含rHBsAg 20 μg和氢氧化铝500 μg),混合均匀,无菌分装,每支1ml。每ml含蝇蛆抗菌肽50μg、rHBsAg 10μg和氢氧化铝250μg。
实施例4 蝇蛆抗菌肽对卵清蛋白受免小鼠免疫反应的佐剂作用
实验方法:清洁级ICR小鼠随机分6组,每组6只。生理盐水对照组:每鼠注射生理盐水(saline)0.2 ml;卵清蛋白(OVA)对照组:每鼠注射含OVA 25 μg的生理盐水0.2 ml;Quil A对照组:每鼠注射含10μg Quil A和25 μg OVA的生理盐水 0.2 ml;蝇蛆抗菌肽实验组:每鼠注射含蝇蛆抗菌肽(5、10、25、50 μg) 和25 μg OVA的生理盐水 0.2 ml。各组皮下注射免疫2次,第一次免疫和第二次免疫间隔14天。二免后14天,采血,分离血清,检测特异性抗体效价;将小鼠断颈处死,无菌条件下取脾,制备脾细胞悬液,进行脾细胞增殖反应和细胞因子分析。
结果表明:蝇蛆抗菌肽能显著提高OVA免疫小鼠血清中OVA特异性IgG、IgG1、IgG2a和 IgG2b抗体效价(图1);增强Con A和LPS诱导的OVA受免小鼠脾细胞增殖反应(图2~图3);促进免疫小鼠脾细胞分泌细胞因子IL-2、INF-γ和IL-10的能力(图4~图6)。说明蝇蛆抗菌肽不仅能促进免疫小鼠对OVA的体液免疫和细胞免疫应答,而且可诱导平衡的Th1/Th2免疫反应,对模式抗原OVA具有显著的佐剂作用。
实施例5 蝇蛆总抗菌肽对禽流感–新城疫重组二联活疫苗的佐剂作用
实验方法:将蝇蛆抗菌肽以生理盐水溶解制成浓度分别为0.5、1.0、2.0和3.0 mg/ml的稀释液,以0.22μm微孔滤膜滤过,备用。将Quil A以生理盐水溶解制成0.1 mg/ml的稀释液。取商品禽流感–新城疫重组二联活疫苗(rL-H5,500羽份/瓶)1瓶加入生理盐水 50 ml,混匀,制成疫苗稀释液。取疫苗稀释液与等量的生理盐水、Quil A稀释液、蝇蛆抗菌肽(MDPF)稀释液,混匀,分别于第1和15天皮下注射免疫小鼠。二免后14天,采血,分离血清,进行H5抗原特异性抗体效价检测;将小鼠断颈处死,无菌条件下取脾,制备脾细胞悬液,进行脾细胞增殖反应、自然杀伤(NK)细胞活性和细胞因子分析。
结果表明:蝇蛆抗菌肽(MDPF)能显著提高禽流感–新城疫重组二联活疫苗(rL-H5)免疫小鼠血清中H5抗原特异性IgG、IgG1、IgG2a和IgG2b抗体滴度(图7);增强Con A 、LPS和H5抗原诱导的受免小鼠脾细胞增殖反应(图8~10)以及NK细胞对K562细胞的杀伤活性(图11);促进免疫小鼠脾细胞分泌细胞因子IL-2、INF-γ和IL-10的能力(图12~14)。说明蝇蛆抗菌肽不仅能促进免疫小鼠对禽流感–新城疫重组二联活疫苗(rL-H5)的体液免疫和细胞免疫应答,而且可同时诱导免疫小鼠对rL-H5疫苗的Th1型和Th2型免疫反应,对禽流感–新城疫重组二联活疫苗具有显著的佐剂作用。
综上所述,本发明的蝇蛆抗菌肽具有显著的免疫佐剂作用,可促进免疫小鼠的细胞免疫和体液免疫应答,诱生机体同时产生平衡的Th1型和Th2型免疫反应,显示出比现有技术中已知的铝胶佐剂更佳的佐剂特性。同时,以该抗菌肽为佐剂的疫苗制备工艺简单、方法简便、质量容易控制、使用方便、可冰冻保存、安全性好。因此,本发明的蝇蛆抗菌肽有望开发成疫苗佐剂。
Claims (6)
1.蝇蛆抗菌肽佐剂,其特征是从蝇蛆中提取、分离的蝇蛆多肽组分。
2.制备权利要求1所述的蝇蛆抗菌肽佐剂的方法,该方法包括以下步骤:
a. 将蝇蛆以质量浓度3%醋酸提取;
b. 醋酸提取液100℃加热1 h,4℃静置,滤过;
c. 滤液浓缩,冷冻干燥,得蝇蛆提取物;
d. 蝇蛆提取物上大孔吸附树脂柱进行色谱纯化,以水和55%乙醇依次洗脱,收集55%乙醇洗脱液,浓缩,冷冻干燥,得蝇蛆抗菌肽。
3.根据权利要求2所述的制备方法,其特征是步骤d中所说的大孔吸附树脂为D101大孔吸附树脂。
4.含蝇蛆抗菌肽佐剂的疫苗制剂,其特征在于包含有免疫有效量的权利要求1所述的蝇蛆抗菌肽佐剂和药物学上可接受的载体。
5.根据权利要求4所述的疫苗制剂,其特征在于蝇蛆抗菌肽佐剂的量为0.01 μg~1g /单剂量。
6.根据权利要求1所述的蝇蛆抗菌肽佐剂在制备疫苗制剂中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410133703.1A CN103908668B (zh) | 2014-04-03 | 2014-04-03 | 蝇蛆抗菌肽佐剂及其制备方法和含该佐剂的疫苗制剂及用途 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410133703.1A CN103908668B (zh) | 2014-04-03 | 2014-04-03 | 蝇蛆抗菌肽佐剂及其制备方法和含该佐剂的疫苗制剂及用途 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN103908668A true CN103908668A (zh) | 2014-07-09 |
CN103908668B CN103908668B (zh) | 2016-06-29 |
Family
ID=51034870
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201410133703.1A Expired - Fee Related CN103908668B (zh) | 2014-04-03 | 2014-04-03 | 蝇蛆抗菌肽佐剂及其制备方法和含该佐剂的疫苗制剂及用途 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN103908668B (zh) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108130810A (zh) * | 2017-12-06 | 2018-06-08 | 王敏 | 一种造纸干强剂的制备方法 |
CN109022522A (zh) * | 2018-08-31 | 2018-12-18 | 杭州洪晟生物技术股份有限公司 | 一种gfp-2小肽及其制备方法和应用 |
CN111057738A (zh) * | 2020-01-02 | 2020-04-24 | 浙江大学 | C2c12细胞在疫苗佐剂机制研究中的用途和研究分析方法 |
CN113521007A (zh) * | 2016-07-01 | 2021-10-22 | 四川大学 | 抗菌肽衍生物在制备免疫佐剂中的用途 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102425918A (zh) * | 2011-12-12 | 2012-04-25 | 舟山市普陀新展望水产饲料有限公司 | 蝇蛆的干燥方法 |
CN102558387A (zh) * | 2010-12-13 | 2012-07-11 | 青岛中仁药业有限公司 | 一种从蝇蛆中提取甲壳素和抗菌肽的方法 |
-
2014
- 2014-04-03 CN CN201410133703.1A patent/CN103908668B/zh not_active Expired - Fee Related
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102558387A (zh) * | 2010-12-13 | 2012-07-11 | 青岛中仁药业有限公司 | 一种从蝇蛆中提取甲壳素和抗菌肽的方法 |
CN102425918A (zh) * | 2011-12-12 | 2012-04-25 | 舟山市普陀新展望水产饲料有限公司 | 蝇蛆的干燥方法 |
Non-Patent Citations (3)
Title |
---|
吴强: "蝇蛆蛋白及其抗菌肽在兽医学上的研究进展", 《黑龙江畜牧兽医》, 31 August 2009 (2009-08-31), pages 21 - 23 * |
过振宇: "大孔吸附树脂对家蝇抗菌肽的吸附与分离纯化", 《天然产物研究与开发》, vol. 20, 31 December 2008 (2008-12-31), pages 969 - 973 * |
陆婕等: "家蝇蛆抗菌肽提取工艺研究", 《昆虫学报》, vol. 50, no. 2, 28 February 2007 (2007-02-28), pages 106 - 107 * |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113521007A (zh) * | 2016-07-01 | 2021-10-22 | 四川大学 | 抗菌肽衍生物在制备免疫佐剂中的用途 |
CN108130810A (zh) * | 2017-12-06 | 2018-06-08 | 王敏 | 一种造纸干强剂的制备方法 |
CN109022522A (zh) * | 2018-08-31 | 2018-12-18 | 杭州洪晟生物技术股份有限公司 | 一种gfp-2小肽及其制备方法和应用 |
CN109022522B (zh) * | 2018-08-31 | 2021-08-17 | 杭州洪晟生物技术股份有限公司 | 一种gfp-2小肽及其制备方法和应用 |
CN111057738A (zh) * | 2020-01-02 | 2020-04-24 | 浙江大学 | C2c12细胞在疫苗佐剂机制研究中的用途和研究分析方法 |
Also Published As
Publication number | Publication date |
---|---|
CN103908668B (zh) | 2016-06-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Wu et al. | Effect of a polysaccharide from Poria cocos on humoral response in mice immunized by H1N1 influenza and HBsAg vaccines | |
Da Hora et al. | Non-toxic derivatives of LT as potent adjuvants | |
Tagliabue et al. | Vaccine adjuvants: the dream becomes real | |
Edelman | Survey of human-use adjuvants | |
CN101402666B (zh) | 具有免疫佐剂作用的皂苷及其制备方法和含该皂苷为佐剂的疫苗制剂及用途 | |
US20120014991A1 (en) | Novel, non-antigenic, mucosal adjuvant formulation which modulates the effects of substances, including vaccine antigens, in contact with mucosal body surfaces | |
CN103908668B (zh) | 蝇蛆抗菌肽佐剂及其制备方法和含该佐剂的疫苗制剂及用途 | |
Song et al. | Ginseng stem–leaf saponins (GSLS) and mineral oil act synergistically to enhance the immune responses to vaccination against foot-and-mouth disease in mice | |
Gupta et al. | Immunological adjuvant effect of Boswellia serrata (BOS 2000) on specific antibody and cellular response to ovalbumin in mice | |
CN102481312A (zh) | 合成的吡喃葡萄糖脂佐剂 | |
JP2006232799A (ja) | ポリ−γ−グルタミン酸含有免疫補強剤組成物 | |
JP6534443B2 (ja) | 竹発酵抽出物の製造方法及び免疫賦活用食品組成物又は免疫賦活剤の製造方法 | |
Xu et al. | The immune protection induced by a serine protease from the Trichinella spiralis adult administered as DNA and protein vaccine | |
Patel | A review on herbal immunoadjuvant. | |
CN103768593A (zh) | 合欢皮皂苷佐剂及其制备方法和含该佐剂的疫苗制剂及用途 | |
JP2018530620A (ja) | 同一の分野の発明を含有する新規アジュバント及びワクチン組成物 | |
AU730230B2 (en) | Novel saponin compositions and uses thereof | |
Weng et al. | Chemical composition and adjuvant properties of the macromolecules from cultivated Cistanche deserticola YC Ma as an immunopotentiator | |
CN101838325B (zh) | 猪用抗原呈递蛋白及其编码基因和应用 | |
CN105169386B (zh) | 一种新型通用型基质疫苗佐剂及其制备方法和用途 | |
CN106692260A (zh) | 一种治疗上呼吸道感染的药物组合物及其应用 | |
Miao et al. | Immuno-Enhancing Effect of Ginsenoside Rh2 Liposomes on Foot-and-Mouth Disease Vaccine | |
CN102085366B (zh) | 一种复合疫苗佐剂 | |
CN106039305B (zh) | 天麻提取物的应用及含有天麻提取物的免疫佐剂和疫苗 | |
CN116036262A (zh) | 一种人参酸性多糖疫苗佐剂、疫苗组合物及其应用 |
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: 20160629 |
|
CF01 | Termination of patent right due to non-payment of annual fee |