CN113476598A - 一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用 - Google Patents
一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用 Download PDFInfo
- Publication number
- CN113476598A CN113476598A CN202110819848.7A CN202110819848A CN113476598A CN 113476598 A CN113476598 A CN 113476598A CN 202110819848 A CN202110819848 A CN 202110819848A CN 113476598 A CN113476598 A CN 113476598A
- Authority
- CN
- China
- Prior art keywords
- novel coronavirus
- vaccine
- nano
- porphyrin
- protein
- 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.)
- Pending
Links
- 229960005486 vaccine Drugs 0.000 title claims abstract description 80
- 241000711573 Coronaviridae Species 0.000 title claims abstract description 66
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 229920000747 poly(lactic acid) Polymers 0.000 claims abstract description 30
- 102000036639 antigens Human genes 0.000 claims abstract description 28
- 108091007433 antigens Proteins 0.000 claims abstract description 28
- 239000004626 polylactic acid Substances 0.000 claims abstract description 27
- 150000002894 organic compounds Chemical class 0.000 claims abstract description 22
- 150000004032 porphyrins Chemical class 0.000 claims abstract description 22
- 230000008685 targeting Effects 0.000 claims abstract description 21
- 150000004033 porphyrin derivatives Chemical class 0.000 claims abstract description 19
- 229920001184 polypeptide Polymers 0.000 claims abstract description 18
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 18
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 18
- XLJKHNWPARRRJB-UHFFFAOYSA-N cobalt(2+) Chemical compound [Co+2] XLJKHNWPARRRJB-UHFFFAOYSA-N 0.000 claims abstract description 17
- 150000002500 ions Chemical class 0.000 claims abstract description 17
- 210000000612 antigen-presenting cell Anatomy 0.000 claims abstract description 13
- 239000012646 vaccine adjuvant Substances 0.000 claims abstract description 12
- 229940124931 vaccine adjuvant Drugs 0.000 claims abstract description 12
- 150000002632 lipids Chemical class 0.000 claims abstract description 10
- 208000001528 Coronaviridae Infections Diseases 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 23
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 16
- 239000002539 nanocarrier Substances 0.000 claims description 16
- 108091005634 SARS-CoV-2 receptor-binding domains Proteins 0.000 claims description 15
- 108090000623 proteins and genes Proteins 0.000 claims description 13
- 102000004169 proteins and genes Human genes 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 8
- 239000011258 core-shell material Substances 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 239000002504 physiological saline solution Substances 0.000 claims description 7
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 6
- 150000001413 amino acids Chemical group 0.000 claims description 4
- 229910001429 cobalt ion Inorganic materials 0.000 claims description 4
- 238000006482 condensation reaction Methods 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 claims description 4
- 238000005199 ultracentrifugation Methods 0.000 claims description 4
- 238000001704 evaporation Methods 0.000 claims description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 3
- 239000000700 radioactive tracer Substances 0.000 claims description 3
- 201000010099 disease Diseases 0.000 claims description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 2
- 230000002163 immunogen Effects 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 108010076039 Polyproteins Proteins 0.000 claims 3
- 239000011807 nanoball Substances 0.000 claims 3
- 230000000890 antigenic effect Effects 0.000 claims 2
- 239000002671 adjuvant Substances 0.000 abstract description 21
- 210000004027 cell Anatomy 0.000 abstract description 10
- 238000006243 chemical reaction Methods 0.000 abstract description 9
- 230000000840 anti-viral effect Effects 0.000 abstract description 3
- 230000005847 immunogenicity Effects 0.000 abstract description 3
- 239000012634 fragment Substances 0.000 description 24
- 241000699666 Mus <mouse, genus> Species 0.000 description 23
- 210000004443 dendritic cell Anatomy 0.000 description 21
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 18
- 238000001514 detection method Methods 0.000 description 13
- 239000002105 nanoparticle Substances 0.000 description 13
- 238000011068 loading method Methods 0.000 description 11
- 210000001165 lymph node Anatomy 0.000 description 11
- 239000000427 antigen Substances 0.000 description 9
- 102100031673 Corneodesmosin Human genes 0.000 description 8
- 101710139375 Corneodesmosin Proteins 0.000 description 8
- 230000002209 hydrophobic effect Effects 0.000 description 8
- 210000002966 serum Anatomy 0.000 description 8
- 239000003814 drug Substances 0.000 description 7
- 230000003053 immunization Effects 0.000 description 7
- 210000004072 lung Anatomy 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 210000001185 bone marrow Anatomy 0.000 description 6
- 229940079593 drug Drugs 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 238000002649 immunization Methods 0.000 description 6
- FDKRLXBXYZKWRZ-UWJYYQICSA-N 3-[(21S,22S)-16-ethenyl-11-ethyl-4-hydroxy-12,17,21,26-tetramethyl-7,23,24,25-tetrazahexacyclo[18.2.1.15,8.110,13.115,18.02,6]hexacosa-1,4,6,8(26),9,11,13(25),14,16,18(24),19-undecaen-22-yl]propanoic acid Chemical group CCC1=C(C2=NC1=CC3=C(C4=C(CC(=C5[C@H]([C@@H](C(=CC6=NC(=C2)C(=C6C)C=C)N5)C)CCC(=O)O)C4=N3)O)C)C FDKRLXBXYZKWRZ-UWJYYQICSA-N 0.000 description 5
- 241000699670 Mus sp. Species 0.000 description 5
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 238000000502 dialysis Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 4
- 229920000469 amphiphilic block copolymer Polymers 0.000 description 4
- 239000006285 cell suspension Substances 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- ZSIAUFGUXNUGDI-UHFFFAOYSA-N hexan-1-ol Chemical compound CCCCCCO ZSIAUFGUXNUGDI-UHFFFAOYSA-N 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 3
- JJTUDXZGHPGLLC-IMJSIDKUSA-N 4511-42-6 Chemical compound C[C@@H]1OC(=O)[C@H](C)OC1=O JJTUDXZGHPGLLC-IMJSIDKUSA-N 0.000 description 3
- 108010088751 Albumins Proteins 0.000 description 3
- 102000009027 Albumins Human genes 0.000 description 3
- 208000025721 COVID-19 Diseases 0.000 description 3
- 208000032376 Lung infection Diseases 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 108090001074 Nucleocapsid Proteins Proteins 0.000 description 3
- 206010035664 Pneumonia Diseases 0.000 description 3
- 229940096437 Protein S Drugs 0.000 description 3
- 210000001744 T-lymphocyte Anatomy 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 239000000969 carrier Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 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 3
- 210000002540 macrophage Anatomy 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 102100035765 Angiotensin-converting enzyme 2 Human genes 0.000 description 2
- 108090000975 Angiotensin-converting enzyme 2 Proteins 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 241001678559 COVID-19 virus Species 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 102100038132 Endogenous retrovirus group K member 6 Pro protein Human genes 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 description 2
- 102100037850 Interferon gamma Human genes 0.000 description 2
- 108010074328 Interferon-gamma Proteins 0.000 description 2
- 102000000588 Interleukin-2 Human genes 0.000 description 2
- 108010002350 Interleukin-2 Proteins 0.000 description 2
- 241000713666 Lentivirus Species 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 description 2
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 230000007969 cellular immunity Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 230000008034 disappearance Effects 0.000 description 2
- 239000002158 endotoxin Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 108010034529 leucyl-lysine Proteins 0.000 description 2
- 229920006008 lipopolysaccharide Polymers 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 230000004807 localization Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 150000003904 phospholipids Chemical class 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 229950010131 puromycin Drugs 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000000108 ultra-filtration Methods 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 102100030988 Angiotensin-converting enzyme Human genes 0.000 description 1
- JUDGRMABQJKRPW-XIADSQHASA-N CCC1=C(/C=C2\N=C(/C(\CC3=O)=C(/[C@@H](CCC(O)=O)[C@@H]4C)\N/C\4=C\C(C(C)=C4C=C)=N/C\4=C4)C3=C\2C)NC/4=C1C Chemical compound CCC1=C(/C=C2\N=C(/C(\CC3=O)=C(/[C@@H](CCC(O)=O)[C@@H]4C)\N/C\4=C\C(C(C)=C4C=C)=N/C\4=C4)C3=C\2C)NC/4=C1C JUDGRMABQJKRPW-XIADSQHASA-N 0.000 description 1
- 108010061994 Coronavirus Spike Glycoprotein Proteins 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 101710091045 Envelope protein Proteins 0.000 description 1
- FKXCBKCOSVIGCT-AVGNSLFASA-N Gln-Lys-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O FKXCBKCOSVIGCT-AVGNSLFASA-N 0.000 description 1
- ITYRYNUZHPNCIK-GUBZILKMSA-N Glu-Ala-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O ITYRYNUZHPNCIK-GUBZILKMSA-N 0.000 description 1
- HGJREIGJLUQBTJ-SZMVWBNQSA-N Glu-Trp-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(C)C)C(O)=O HGJREIGJLUQBTJ-SZMVWBNQSA-N 0.000 description 1
- 108060003393 Granulin Proteins 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 102100022297 Integrin alpha-X Human genes 0.000 description 1
- DLCOFDAHNMMQPP-SRVKXCTJSA-N Leu-Asp-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O DLCOFDAHNMMQPP-SRVKXCTJSA-N 0.000 description 1
- XXXXOVFBXRERQL-ULQDDVLXSA-N Leu-Pro-Phe Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 XXXXOVFBXRERQL-ULQDDVLXSA-N 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical class OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 1
- 208000034530 PLAA-associated neurodevelopmental disease Diseases 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- MPGJIHFJCXTVEX-KKUMJFAQSA-N Phe-Arg-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O MPGJIHFJCXTVEX-KKUMJFAQSA-N 0.000 description 1
- 229920001244 Poly(D,L-lactide) Polymers 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 101710188315 Protein X Proteins 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 101710198474 Spike protein Proteins 0.000 description 1
- 229940124614 TLR 8 agonist Drugs 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 238000005411 Van der Waals force Methods 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000004721 adaptive immunity Effects 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000020411 cell activation Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 230000009920 chelation Effects 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 230000001112 coagulating effect Effects 0.000 description 1
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000011577 humanized mouse model Methods 0.000 description 1
- 230000004727 humoral immunity Effects 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 231100000225 lethality Toxicity 0.000 description 1
- 238000010859 live-cell imaging Methods 0.000 description 1
- 210000001365 lymphatic vessel Anatomy 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 229910001453 nickel ion Inorganic materials 0.000 description 1
- 125000004433 nitrogen atom Chemical group N* 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000002823 phage display Methods 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 208000023504 respiratory system disease Diseases 0.000 description 1
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 1
- 238000007151 ring opening polymerisation reaction Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- KSBAEPSJVUENNK-UHFFFAOYSA-L tin(ii) 2-ethylhexanoate Chemical compound [Sn+2].CCCCC(CC)C([O-])=O.CCCCC(CC)C([O-])=O KSBAEPSJVUENNK-UHFFFAOYSA-L 0.000 description 1
- 229940044616 toll-like receptor 7 agonist Drugs 0.000 description 1
- 229940044655 toll-like receptor 9 agonist Drugs 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 108010015666 tryptophyl-leucyl-glutamic acid Proteins 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000008478 viral entry into host cell Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/52—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an inorganic compound, e.g. an inorganic ion that is complexed with the active ingredient
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/593—Polyesters, e.g. PLGA or polylactide-co-glycolide
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6921—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6925—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a microcapsule, nanocapsule, microbubble or nanobubble
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0036—Porphyrins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0054—Macromolecular compounds, i.e. oligomers, polymers, dendrimers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G63/00—Macromolecular compounds obtained by reactions forming a carboxylic ester link in the main chain of the macromolecule
- C08G63/91—Polymers modified by chemical after-treatment
- C08G63/912—Polymers modified by chemical after-treatment derived from hydroxycarboxylic acids
-
- 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
- A61K2039/55588—Adjuvants of undefined constitution
-
- 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
- A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
- A61K2039/572—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 cytotoxic response
-
- 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
- A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
- A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
-
- 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
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
-
- 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
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
- A61K2039/6031—Proteins
-
- 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
- A61K2039/62—Medicinal preparations containing antigens or antibodies characterised by the link between antigen and carrier
- A61K2039/622—Medicinal preparations containing antigens or antibodies characterised by the link between antigen and carrier non-covalent binding
-
- 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
- C12N2770/20011—Coronaviridae
- C12N2770/20034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nanotechnology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Virology (AREA)
- Microbiology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Mycology (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Biomedical Technology (AREA)
- Pulmonology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Engineering & Computer Science (AREA)
- Communicable Diseases (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Oncology (AREA)
- Medical Informatics (AREA)
- Inorganic Chemistry (AREA)
- Polymers & Plastics (AREA)
- Physics & Mathematics (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- General Physics & Mathematics (AREA)
- Manufacturing & Machinery (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用,所述纳米疫苗包括:由聚乳酸、卟啉或卟啉衍生物以及Co2+离子结合物自组装的有机化合物;新型冠状病毒抗原蛋白;疫苗佐剂和脂质。本发明合成的有机化合物,其核心包裹有佐剂,表面高效装载有新型冠状病毒抗原蛋白,实现了新型冠状病毒抗原蛋白和疫苗佐剂共同递送的纳米疫苗体系,不仅可最大限度发挥新型冠状病毒重组亚蛋白的免疫原性,还可通过荧光分子示踪其在生物体内分布情况。此外,本发明的纳米疫苗还连接了可特异性靶向抗原提呈细胞的多肽,促进纳米疫苗被DC细胞大量摄取,促进抗病毒反应。同时本发明的制备方法简单高效,为有效预防新型冠状病毒感染提供了一个全新途径。
Description
技术领域
本发明属于生物医药领域,具体涉及一种新型冠状病毒亚蛋白纳米疫苗及 其制备方法和应用。
背景技术
新型冠状病毒肺炎(COVID-19)是由新型冠状病毒(SARS-CoV-2)引起 的一种呼吸系统疾病,其临床表现从轻症的流感类症状到重症危机生命的肺炎, 具备非常高的传染性和致死率。其主要结构包括刺突蛋白(S,spike)、包膜蛋 白(E,envelope)、膜蛋白/基质蛋白(M,membrane/matrix)和核衣壳蛋白(N, uncleocapsid)。当宿主感染新型冠状病毒后,S蛋白会被蛋白酶切割分成S1亚 基和S2亚基,S1亚基的受体结合域(RBD)会通过与宿主细胞表面的血管紧 张素转化酶2(Angiotensin-converting enzyme 2,ACE2)结合,引起发烧以及肺 部感染等临床表现。针对S蛋白的中和抗体可以阻断病毒侵入宿主细胞。因此, 接种针对S蛋白开发的新型冠状病毒疫苗的是改善疫情蔓延的有效方法。
近年来,致力于实现抗原和佐剂共同递送的纳米疫苗平台开发迅速,脂质 体,白蛋白以及一些通过FDA的两亲性嵌段共聚物是近些年用于递送平台的纳 米载体,脂质体是指由两亲型磷脂分子为基础材料,加入胆固醇等其他辅料, 在水中由于两亲性磷脂疏水段聚集在一起,亲水端暴露在水中原因从而自组装 形成中间疏水两端亲水的双分子层结构囊泡;白蛋白是血浆中最丰富的蛋白, 使用白蛋白作为难溶性药物的载体制备药物递送纳米粒子,会使其具有低生物 毒性,低免疫反应,可生物降解和提升半衰期等优点,白蛋白与疏水小分子药 物之间的结合力主要为范德华力和氢键作用力,疏水类小分子药物主要装载牛 血清白蛋白的IIA区域,能够显著的提升药物在血浆中的溶解度;两亲性嵌段 共聚物也是主要依靠自身的疏水结构域通过亲疏水作用力装载疏水性小分子药 物,例如PLGA,凭借着其优异的生物相容性,低生物毒性和在生物体内良好的 降解性使其通过美国食品药品管理局的认证并被正式作为药用辅料收录进美国 药典,并被批准用于多种药物的体内递送系统,但是上述载体均很难实现新型 冠状病毒S蛋白的装载,这是由于新冠S蛋白的分子量较大(140KD),具有 较大的空间位阻效应,无法有效大容量装载于纳米体系中;并且S蛋白的抗原 的空间效应使之只能通过吸附作用或者油包水装载到纳米粒子内核,上述两种 装载方式不仅效率低下,并且更容易释放,难以稳定保存。
发明内容
本发明的目的在于提供一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和 应用,通过将聚乳酸与卟啉类物质偶联,并螯合有可与组氨酸标签连接的Co2+离子,形成由聚乳酸、卟啉或卟啉衍生物以及Co2+离子结合物自组装的有机化 合物,该有机化合物的核心中包裹有佐剂,核壳表面包裹有脂质,表面还高效 装载有新型冠状病毒抗原蛋白,从而实现了新型冠状病毒抗原蛋白和疫苗佐剂 共同递送的纳米疫苗体系,该体系不仅可最大限度发挥新型冠状病毒重组亚蛋 白的免疫原性,同时还可通过荧光分子示踪其在生物体内分布情况;进一步还 通过设计并装载有可特异性靶向抗原提呈细胞的多肽,从而更有效促进机体的 抗病毒反应。
为实现上述目的,本发明采用的技术方案是:
本发明提供了一种新型冠状病毒亚蛋白纳米疫苗,所述纳米疫苗包括:由 聚乳酸、卟啉或卟啉衍生物以及Co2+离子结合物自组装的有机化合物;新型冠 状病毒抗原蛋白;疫苗佐剂和脂质。
进一步地,所述纳米疫苗中由聚乳酸、卟啉或卟啉衍生物以及Co2+离子结 合物自组装的有机化合物为核壳结构,核内为疫苗佐剂,核壳上包裹有脂质, 核壳表面负载有新型冠状病毒抗原蛋白。
进一步地,所述核壳表面还负载有靶向抗原提呈细胞的靶向多肽,其中所 述靶向多肽的氨基酸序列如SEQ ID NO:1所示。其中所述抗原提呈细胞包括: 树突状细胞和巨噬细胞,并且该靶向多肽通过噬菌体展示技术获得。
进一步地,所述新型冠状病毒抗原蛋白和靶向多肽分别通过组氨酸标签与 有机化合物连接。
进一步地,所述新型冠状病毒抗原蛋白为新型冠状病毒的RBD蛋白。
进一步地,所述卟啉衍生物为只含有一个羧基基团的卟啉衍生物。
进一步地,所述卟啉衍生物为焦脱镁叶绿酸-α。
进一步地,所述疫苗佐剂可选自:TLR7激动剂、TLR8激动剂、TLR9激动 剂、QS-21佐剂。
进一步地,所述聚乳酸的分子量大小优选为1500-3000。
进一步地,所述脂质为PEG-磷脂,优选为PEG-DSPE,更优选为 DSPE-PEG2000。
进一步地,所述有机化合物的结构式为:
本发明还提供了上述新型冠状病毒亚蛋白纳米疫苗的制备方法,包括:
步骤1、将聚乳酸与卟啉或卟啉衍生物反应,使聚乳酸末端的羟基与卟啉或 卟啉衍生物的羧基进行缩合反应,使卟啉或卟啉衍生物键合到聚乳酸聚合物链 的末端;
步骤2、向步骤1的产物中加入Co2+离子,使Co2+离子嵌入卟啉环中,得 到由聚乳酸、卟啉或卟啉衍生物以及Co2+离子结合物自组装的有机化合物;
步骤3、将步骤2制得的有机化合物、脂质和疫苗佐剂混合溶解,滴入生理 盐水中,蒸发有机溶剂获得含有钴离子卟啉环的均匀分布于生理盐水中的纳米 载体溶液;
步骤4、将连接有组氨酸标签的新型冠状病毒抗原蛋白,与步骤3制得的纳 米载体溶液孵育过夜,超速离心获得结合有新型冠状病毒抗原蛋白的纳米疫苗。
进一步的,所述制备方法中还包括:将含有组氨酸标签的新型冠状病毒抗 原蛋白以及特异性靶向多肽同时与纳米载体溶液孵育过夜,然后超速离心获得 纳米疫苗。
进一步的,所述步骤1中,向焦脱镁叶绿酸-α中加入聚乳酸,然后加入1-(3- 二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和4-二甲氨基吡啶,冰浴反应至检测到起 始醇消失,然后将反应液经萃取、洗涤干燥,并除去溶剂后得到产物PLA-焦脱 镁叶绿酸。
本发明还提供了上述新型冠状病毒亚蛋白纳米疫苗在制备与新型冠状病毒 感染相关疾病的免疫原性组合物中的应用。
本发明还提供了上述新型冠状病毒亚蛋白纳米疫苗在制备纳米示踪剂中的 应用。
与现有技术相比,本发明的有益效果是:本发明通过将聚乳酸与卟啉类物 质偶联并螯合有Co2+离子,形成了由聚乳酸、卟啉或卟啉衍生物以及Co2+离子 结合物自组装的有机化合物,该有机化合物通过Co2+离子与组氨酸标签连接, 实现了高效装载新型冠状病毒抗原蛋白;并利用化学偶联的卟啉类物质本身可 以产生荧光的功能,示踪纳米颗粒的在体分布;同时还在核心包裹疫苗有佐剂, 核壳表面包裹有脂质,从而实现了新型冠状病毒抗原蛋白和疫苗佐剂共同递送 的纳米疫苗体系。进一步地,本发明还设计并在化合物表面装载有可靶向抗原 提呈细胞的多肽,从而促进纳米疫苗被DC细胞大量摄取,更有效促进机体的抗 病毒反应。即本发明提供了一种兼具高效装载抗原、同时携载佐剂和抗原、示 踪纳米疫苗分布并特异性高效靶向DC细胞功能的新型冠状病毒亚蛋白纳米疫 苗,并且其制备方法简单高效,为有效预防新型冠状病毒感染、新型冠状病毒 亚蛋白纳米疫苗的制备提供了一个全新高效的途径与选择。
附图说明
图1为本发明实施例1中制备的纳米疫苗NPQ-RBD结构示意图;
图2为本发明实施例1中特异性靶向抗原提呈细胞的多肽的获取方式;
图3为本发明实施例1中制备的纳米疫苗NPQ-RBD-AP结构示意图;
图4为本发明实施例1中纳米疫苗的稳定性检测结果;
图5为本发明实施例2中纳米疫苗对小鼠骨髓来源树突状细胞的体外靶向 性检测结果;
图6为本发明实施例2中纳米疫苗对小鼠骨髓来源树突状细胞的激活效应 检测结果;
图7为本发明实施例2中纳米疫苗靶向淋巴结和淋巴结中的抗原提呈细胞 的检测结果;
图8为本发明实施例2中纳米疫苗诱导动物血清产生抗RBD片段的抗体效 价检测结果;
图9为本发明实施例2中纳米疫苗NPQ-RBD诱导动物血清产生抗RBD片 段的抗体效价随时间变化的检测结果;
图10为本发明实施例2中纳米疫苗诱导机体产生针对RBD片段的细胞免 疫效应检测结果;
图11为本发明实施例2中纳米疫苗对hACE2人源性小鼠抵抗新型冠状病 毒COVID-19导致的肺部感染的检测结果。
具体实施方式
下面将结合本发明中的实施例,对本发明的技术方案进行清楚、完整地描 述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。 基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动条件下所 获得的所有其它实施例,都属于本发明保护的范围。
实施例1新型冠状病毒亚蛋白纳米疫苗的制备
1、聚乳酸PLA的合成
具体操作步骤为:
于手套箱中称取0.2500g正己醇置于干燥的安瓿瓶中,加入L-LA,D-LA 各2.1180g,加入0.31mL预先配置好的浓度为0.02g/mL的Sn(Oct)2溶液,最后 加入21.0mL干燥的甲苯。聚合反应在130℃、N2环境中反应2天。溶液用200 mL冰乙醚沉降,得到的产物室温下干燥24h。然后产物用N,N-二甲基甲酰胺 (DMF)溶解,通过透析袋(MWCO=1000D)在DMF中透析48h除去少量未 反应的单体,转移至去离子水中透析去除有机溶剂,经冷冻干燥得最终产物。
1H NMR各个峰的成功归属及峰面积的计算证明了该聚合物成功合成。目 前已知的PLA分子量可从几百到几万,其自身分子量越大疏水性越强,与辅料 的分子量相差越大,其装载能力越弱。即通过调节投料比可制备得到不同的分 子量的PLA,而基于不同分子量的PLA制备的纳米疫苗对于目标蛋白的装载能 力有所不同。经优选,本发明选用分子量1500-3000的PLA以最大程度实现新 型冠状病毒抗原蛋白RBD蛋白的装载。
2、PLA与卟啉类物质共价结合
本实施例以卟啉类物质:焦脱镁叶绿酸-α为例,将上述制得的PLA与焦脱 镁叶绿酸-α反应,通过PLA末端的羟基与焦脱镁叶绿酸-α的羧基之间的缩合反 应,使焦脱镁叶绿酸-α键合到PLA聚合物链的末端,合成路径如下:
具体步骤为:
向焦脱镁叶绿酸(53.4mg)的无水二氯甲烷溶液中加入162.3mg PLA,然 后加入38.6mg的1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)、49.8mg 的4-二甲氨基吡啶,于冰浴中反应24h,TCL点板检测起始醇消失后,将反应 液倒入水中,用二氯甲烷多次萃取产物,合并二氯甲烷相,然后依次用 4%NaHCO3水溶液洗涤、饱和食盐水洗涤,无水硫酸镁干燥12h。抽滤得产物 的二氯甲烷溶液,然后通过旋转蒸发仪除去溶剂得到粗产物,通过透析袋 (MWCO=1000D)透析48h除去少量杂质,然后冷冻干燥得到产物PLA-焦脱 镁叶绿酸。所有反应过程避光进行。
1H NMR中Ppa及PDLLA的峰成功归属、反应物Ppa中羧基峰的消失证 明该缩合反应成功进行。紫外-可见吸收光谱(UV-vis)显示该材料与Ppa有相 近的吸收峰位置。
3、Co2+离子的配位螯合
通过在卟啉环中嵌入金属离子,利用金属离子实现与带有组氨酸标签的目 的抗原的结合,本发明经优选发现在所有能够结合组氨酸标签的二价金属离子 中,二价Co2+离子的结合效率最高,因此选用Co2+嵌合到卟啉环中,用于结合 含有组氨酸标签的抗原,Co2+离子可以在分子骨架中心位置于N原子以配位方 式结合,合成路径如下:
具体步骤为:
向上述制得的PLA-焦脱镁叶绿酸(50mg)的二氯甲烷溶液中(10.0mL) 加入氯化钴的饱和甲醇溶液(1.0mL)并搅拌12h。旋转蒸发仪除去溶剂,然后 用2.0mL二氯甲烷溶解,依次用4%NaHCO3水溶液洗涤、饱和食盐水洗涤, 无水硫酸镁干燥,过滤,通过旋转蒸发仪除去溶剂得到产物,即结合钴离子的 卟啉环的PLA有机化合物。UV-vis显示Co2+配位后,吸收峰位置发生红移。
4、RBD蛋白的纯化
RBD片段是新型冠状病毒结合人源性肺上皮细胞ACE2受体的关键结构 域,也是目前常规被用来制备疫苗的最有效的区域。分别构建胞外分泌的和在 细胞中表达的含有组氨酸标签的RBD片段慢病毒质粒,获得能够感染细胞的含 有嘌呤霉素抗性的慢病毒,转染CHO细胞,使用嘌呤霉素筛选获得表达RBD 蛋白片段的CHO细胞系。扩大培养该细胞系,获得上清液或者细胞悬浮液;如 果是细胞悬浮液,通过添加PBS溶液结合超声,获得含有全细胞蛋白的PBS上 清液。使用含有镍离子柱的纯化柱子获得仅表达组氨酸标签的RBD蛋白,接着在生理盐水溶液中进行透析,获得最终所需的RBD蛋白。
5、RBD蛋白的装载
相比较于其它佐剂而言,皂苷佐剂因为低剂量便可以同时引发体液免疫和 细胞免疫效应,被认为是目前最有前景的佐剂,已经被用于肿瘤疫苗和病毒疫 苗的相关临床实验研发,并且由于佐剂的亲疏水特性,能够与纳米体系完美结 合,因此本实施例选用了QS-21作为疫苗佐剂,并选用DSPE-PEG2000磷脂作 为纳米体系。本发明研究发现通过调整DSPE-PEG2000和PLA的含量比例,可 改变该纳米体系结合表达组氨酸标签的蛋白的能力,装载效果最优的体系如下:
用1mL四氢呋喃溶解含有上述有机化合物、DSPE-PEG2000磷脂以及QS-21 佐剂,其中所述有机化合物、DSPE-PEG2000磷脂以及QS-21佐剂的质量比为: 1:10:10。溶解后的溶液逐滴滴入温度为70℃的生理盐水中,旋转蒸发有机溶剂, 获得含有钴离子卟啉环的均匀分布于生理盐水中的纳米载体NPQ溶液;
10KD超滤离心管3000rpm离心获取高浓度含量的包含纳米载体的生理盐 水溶液;将过量的RBD片段与含有纳米载体NPQ的溶液在4℃条件下孵育过夜, 然后通过超速离心获得结合有RBD片段和QS-21的纳米疫苗(NPQ-RBD), 其结构示意图如图1所示。
6、RBD蛋白和靶向多肽的共同装载
淋巴结树突状细胞(DC)是机体重要的抗原提呈细胞,为提高纳米疫苗促 进机体的抗病毒反应,进一步通过噬菌体技术筛选并且合成了一个可特异性靶 向抗原提呈细胞的多肽,其氨基酸序列为:LDLFRELPFEWLEALKQKLK(SEQ ID NO.1所示),将其与RBD蛋白分别通过组氨酸标签与有机化合物连接,并 装载于纳米疫苗表面,具体操作步骤为:
按上述方法获得纳米载体NPQ溶液,10KD超滤离心管3000rpm离心获取 高浓度含量的包含纳米载体的生理盐水溶液;将过量的RBD片段、含有组氨酸 标签的靶向多肽与含有纳米载体的NPQ溶液在4℃条件下孵育过夜,然后通过 超速离心获得结合有RBD片段、靶向多肽和QS-21的纳米疫苗(NPQ-RBD-AP), 其结构示意图如图3所示。
7、不同分子量蛋白与NPQ载体的结合效率分析
为验证本发明优选制得的NPQ载体对于RBD蛋白片段具有高效的结合效 率,分别选取了不同分子量的蛋白质:His-IL-2(含有6个组氨酸标签的白细胞 介素-2)、His-RBD(含有6个组氨酸标签的RBD蛋白)、His-BSA(含有6 个组氨酸标签的牛血清白蛋白)、His-spike protein(含有6个组氨酸标签的新型 冠状病毒Spike的蛋白)作为研究对象;在相等浓度的纳米载体中,加入相同浓 度(50nmol)的上述蛋白,室温反应30分钟后,进行HPLC鉴定,分析游离的 蛋白的含量,并根据公式:加入的总蛋白含量-游离蛋白含量/总蛋白含量*100%, 计算不同分子量蛋白与NPQ载体的结合效率,结果如表1所示:
表1不同分子量与NPQ载体的结合效率
蛋白名称 | 分子量 | 摩尔质量 | 与NPQ结合效率 |
His-IL-2 | 15KD | 50nmol | 72% |
His-RBD | 26.1KD | 50nmol | 92% |
His-BSA | 66.5KD | 50nmol | 75% |
His-spike protein | 190KD | 50nmol | 62% |
结果显示,本发明制备得到的纳米载体NPQ对于RBD蛋白具有最高的装 载效率,其与NPQ的结合效率高达92%。
实施例2纳米疫苗NPQ-RBD和NPQ-RBD-AP的效果验证
1、纳米疫苗的特性表征
采用实施例1所述的方法分别制备得到了纳米疫苗NPQ-RBD和 NPQ-RBD-AP,经检测其粒径为100-200nm之间,该区间大小的纳米颗粒可有 效穿透淋巴管被引流至淋巴结。
为验证携载RBD片段的纳米疫苗NPQ-RBD和NPQ-RBD-AP的稳定性, 使用FITC分子标记了纳米NPQ-RBD上的氨基酸片段;将该纳米至于4℃生理 盐水中透析,每天收集部分溶液,使用酶标仪检测纳米中的相对FITC荧光强度, 并设置不含佐剂的纳米颗粒NP-RBD对照组,以及仅注射RBD抗原的组别,检 测结果如图4所示。
结果显示,本发明制备得到的纳米疫苗NPQ-RBD和NPQ-RBD-AP具有很 好的稳定性,均可稳定一个星期以上,并仍然携带超过70%的RBD片段;
2、纳米疫苗体外靶向和激活小鼠骨髓来源树突状细胞(BMDC)
于无菌条件下提取雄性7-8周小鼠的骨髓,在体外用20ug/mL GM-CSF刺 激7天后,即可获得小鼠骨髓来源树突状细胞BMDC,将BMDC置于共聚焦小 皿中进行培养,FITC染色NPQ-RBD和NPQ-RBD-AP,使用20x物镜拍摄BMDC 摄取纳米的情况,检测结果如图5所示。
结果显示,NPQ-RBD和NPQ-RBD-AP均可被树突状细胞摄取,其中携载 有靶向抗原提呈细胞的多肽的纳米疫苗可被树突状细胞高效摄取。
将BMDC加到六孔板中,5×105/孔,向每孔中加入NPQ-RBD和NPQ-RBD-AP悬液100μL。继续在培养箱中培养24h后,通过流式细胞仪检测 DC细胞活化的标志物CD80、CD86的表达,并以未处理的DC细胞、不含佐剂 和RBD抗原的有机纳米颗粒NP、佐剂QS21以及非特异性活化DC细胞的细菌 脂多糖LPS分别处理DC细胞作为对照,结果如图6所示。
结果显示,相较于未处理的DC细胞,NPQ-RBD和NPQ-RBD-AP处理后 的DC细胞中CD80、CD86的表达显著升高,即证明NPQ-RBD和NPQ-RBD-AP 纳米疫苗可体外能显著激活小鼠骨髓来源树突状细胞(BMDC);此外,该实验结 果也证实了纳米载体NP本身也可以在一定程度上活化DC细胞。
3、纳米疫苗靶向淋巴结和淋巴结中的抗原提呈细胞
使用罗丹明标记纳米疫苗NPQ-RBD-AP上的RBD片段,然后向小鼠的尾 根部注射100μL上述溶液;24h后使用小动物活体成像检测该纳米疫苗到达淋 巴结的程度,检测结果如图7所示;接着将剥离的淋巴结进行冰冻切片以及免 疫荧光操作,使用CD11c抗体标记DC细胞,使用F4-80抗体标记巨噬细胞, 使用激光共聚焦显微镜观察淋巴结中纳米的定位,定位检测结果如图7所示。 结果所示,所述纳米疫苗NPQ-RBD-AP可有效到达引流淋巴结部位,并且能够 特异性富集于抗原提呈细胞区域,如DC细胞和巨噬细胞。
4、纳米疫苗诱导动物血清中产生抗RBD片段的抗体效价检测
在C57小鼠尾根部皮下分别注射100μL携带有RBD片段的纳米疫苗 NPQ-RBD和NPQ-RBD-AP,共免疫两次,于第二次免疫后的第8天和第18天 检测小鼠血清中针对RBD片段的抗体效价,并分别以注射PBS、佐剂QS-21、 不含载体的RBD片段、不含佐剂的纳米颗粒NP-RBD作为对照。
小鼠血清的获取:采用眼眶静脉丛取血的方法,每次获取小鼠血液200-300 μL,常温下凝固半个小时,然后4000rpm离心30分钟,获取小鼠血清;使用 检测RBD抗体的ELisa试剂盒检测小鼠血清中含有的抗RBD片段的IgG抗体 的效价,其中不同处理组的检测结果如图8所示,纳米疫苗NPQ-RBD的抗体效 价随时间变化的检测结果如图9所示。
结果显示,采用本发明制备得到的纳米疫苗可有效诱导RBD区域的抗体产 生,并且产生的抗体效价为对照组的3000倍以上,即证明了该疫苗具有预防新 型冠状病毒的潜力,同时该抗体效价可以在小鼠血清中存在至少10天以上,证 明了该纳米疫苗产生抗体的持久性。
5、纳米疫苗诱导机体产生针对RBD片段的细胞免疫效应
在C57小鼠尾根部皮下注射100μL携带有RBD片段的纳米疫苗NPQ-RBD 和NPQ-RBD-AP,并分别以注射PBS、佐剂QS-21、不含载体的RBD片段、不 含佐剂的纳米颗粒NP-RBD作为对照。共免疫两次,在免疫第二次的一个月后, 劲椎脱臼处死小鼠,提取小鼠脾脏和淋巴结细胞,ACK裂解红细胞,制成单细 胞悬液。加入RBD蛋白片段再次刺激T细胞,使用流式技术检测细胞悬液中的 CD8阳性T细胞中,表达IL-2、TNF-α和IFN-γ等细胞因子的比例,同时检测 CD4阳性T细胞中表达IFN-γ的细胞比例,用于判断小鼠适应性免疫的生成。 检测结果如图10所示。
结果显示,本发明制备得到的纳米疫苗NPQ-RBD和NPQ-RBD-AP均具有 很强的诱导机体产生针对RBD片段的细胞免疫的功效。
6、纳米疫苗抵抗新型冠状病毒COVID-19导致的肺部感染
使用hACE2人源性小鼠,在该小鼠尾根部皮下注射100μL携带有RBD片 段的纳米疫苗NPQ-RBD和NPQ-RBD-AP,并分别以注射PBS、佐剂QS-21、 不含载体的RBD片段、不含佐剂的纳米颗粒NP-RBD作为对照。共免疫两次; 在第二次免疫后的两周,对人源性hACE2小鼠进行攻毒实验,即使用COVID-19 病毒感染该小鼠,在感染小鼠后的一周,使用PBS灌洗小鼠肺部,收集肺灌洗 液,检测小鼠肺部感染新型冠状病毒情况,并通过检测新型冠状病毒mRNA来 判定小鼠感染情况;将小鼠劲椎处死,获取小鼠肺部,并进行HE染色,评判小 鼠肺部炎症情况,检测结果如图11所示,其中图11-B为RNA拷贝数检测结果,11-C为抗体效价检测结果,11-D为肺部炎症观测结果。
结果显示,小鼠肺部无明显病变,即本发明制备得到的纳米疫苗NPQ-RBD 和NPQ-RBD-AP可有效预防hACE2人源性小鼠感染新型冠状病毒。
以上所述,仅为本发明较佳的具体实施方式,但本发明的保护范围并不局 限于此,任何熟悉本技术领域的技术人员在本发明揭露的技术范围内,可轻易 想到的变化或替换,都应涵盖在本发明的保护范围之内。
序列表
<110> 武汉圣润生物科技有限公司
<120> 一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Leu Asp Leu Phe Arg Glu Leu Pro Phe Glu Trp Leu Glu Ala Leu Lys
1 5 10 15
Gln Lys Leu Lys
20
Claims (10)
1.一种新型冠状病毒亚蛋白纳米疫苗,其特征在于,所述纳米疫苗包括:由聚乳酸、卟啉或卟啉衍生物以及Co2+离子结合物自组装的有机化合物;新型冠状病毒抗原蛋白;疫苗佐剂和脂质。
2.根据权利要求1所述的新型冠状病毒亚蛋白纳米疫苗,其特征在于,所述纳米疫苗中由聚乳酸、卟啉或卟啉衍生物以及Co2+离子结合物自组装的有机化合物为核壳结构,核内为疫苗佐剂,核壳上包裹有脂质,核壳表面负载有新型冠状病毒抗原蛋白。
3.根据权利要求2所述的新型冠状病毒亚蛋白纳米疫苗,其特征在于,所述核壳表面还负载有靶向抗原提呈细胞的靶向多肽,其中所述靶向多肽的氨基酸序列如SEQ ID NO:1所示。
4.根据权利要求3所述的新型冠状病毒亚蛋白纳米疫苗,其特征在于,所述新型冠状病毒抗原蛋白和靶向多肽分别通过组氨酸标签与有机化合物连接。
5.根据权利要求1所述的新型冠状病毒亚蛋白纳米疫苗,其特征在于,所述新型冠状病毒抗原蛋白为新型冠状病毒的RBD蛋白。
6.根据权利要求1所述的新型冠状病毒亚蛋白纳米疫苗,其特征在于,所述卟啉衍生物为只含有一个羧基基团的卟啉衍生物。
8.如权利要求1-7任一项所述的新型冠状病毒亚蛋白纳米疫苗的制备方法,其特征在于,所述方法包括:
步骤1、将聚乳酸与卟啉或卟啉衍生物反应,使聚乳酸末端的羟基与卟啉或卟啉衍生物的羧基进行缩合反应,使卟啉或卟啉衍生物键合到聚乳酸聚合物链的末端;
步骤2、向步骤1的产物中加入Co2+离子,使Co2+离子嵌入卟啉环中,得到由聚乳酸、卟啉或卟啉衍生物以及Co2+离子结合物自组装的有机化合物;
步骤3、将步骤2制得的有机化合物、脂质和疫苗佐剂混合溶解,滴入生理盐水中,蒸发有机溶剂获得含有钴离子卟啉环的均匀分布于生理盐水中的纳米载体溶液;
步骤4、将连接有组氨酸标签的新型冠状病毒抗原蛋白,与步骤3制得的纳米载体溶液孵育过夜,超速离心获得结合有新型冠状病毒抗原蛋白的纳米疫苗。
9.如权利要求1-7任一项所述的新型冠状病毒亚蛋白纳米疫苗在制备与新型冠状病毒感染相关疾病的免疫原性组合物中的应用。
10.如权利要求1-7任一项所述的新型冠状病毒亚蛋白纳米疫苗在制备纳米示踪剂中的应用。
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110819848.7A CN113476598A (zh) | 2021-07-20 | 2021-07-20 | 一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用 |
PCT/CN2021/111511 WO2023000403A1 (zh) | 2021-07-20 | 2021-08-09 | 一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110819848.7A CN113476598A (zh) | 2021-07-20 | 2021-07-20 | 一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN113476598A true CN113476598A (zh) | 2021-10-08 |
Family
ID=77942504
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110819848.7A Pending CN113476598A (zh) | 2021-07-20 | 2021-07-20 | 一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN113476598A (zh) |
WO (1) | WO2023000403A1 (zh) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114344457A (zh) * | 2021-11-13 | 2022-04-15 | 暨南大学 | 新型冠状病毒蛋白质抗原纳米疫苗及其制备方法和应用 |
WO2023000403A1 (zh) * | 2021-07-20 | 2023-01-26 | 武汉圣润生物科技有限公司 | 一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用 |
CN115969969A (zh) * | 2023-02-13 | 2023-04-18 | 中国科学院长春应用化学研究所 | 一种仿病毒结构纳米颗粒疫苗及其制备方法和应用 |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106924731A (zh) * | 2015-12-30 | 2017-07-07 | 复旦大学 | 一种基于光动力疗法和化学疗法的联合肿瘤靶向治疗系统 |
CN107708671A (zh) * | 2015-04-16 | 2018-02-16 | 纽约州立大学研究基金会 | 包含钴卟啉‑磷脂缀合物和聚组氨酸标签的纳米结构 |
CN109464395A (zh) * | 2019-01-03 | 2019-03-15 | 中国科学院过程工程研究所 | 一种水包油包凝胶乳液及其制备方法和应用 |
CN111217917A (zh) * | 2020-02-26 | 2020-06-02 | 康希诺生物股份公司 | 一种新型冠状病毒SARS-CoV-2疫苗及其制备方法 |
CN111560074A (zh) * | 2020-03-20 | 2020-08-21 | 中山大学 | 一种基于幽门螺旋杆菌铁蛋白的新型冠状病毒s蛋白单区域亚单位纳米疫苗 |
CN112569207A (zh) * | 2019-09-30 | 2021-03-30 | 复旦大学 | 一种载脂蛋白修饰的仿生纳米肿瘤疫苗及其制备方法和用途 |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103127500B (zh) * | 2013-02-07 | 2015-05-20 | 中山大学 | 卟啉色素作为免疫佐剂和疫苗的用途 |
EP4023249A1 (en) * | 2014-04-23 | 2022-07-06 | ModernaTX, Inc. | Nucleic acid vaccines |
CN104277094A (zh) * | 2014-07-04 | 2015-01-14 | 文康医疗技术(深圳)有限公司 | Dc靶向肽及其应用 |
CN104387453A (zh) * | 2014-12-08 | 2015-03-04 | 深圳市同康生物医药有限公司 | 树突状细胞靶向肽及编码基因及应用 |
JP6757061B2 (ja) * | 2016-05-09 | 2020-09-16 | 東京都公立大学法人 | ドラッグデリバリー用脂質単層被覆型ナノ粒子 |
CN113476598A (zh) * | 2021-07-20 | 2021-10-08 | 武汉圣润生物科技有限公司 | 一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用 |
-
2021
- 2021-07-20 CN CN202110819848.7A patent/CN113476598A/zh active Pending
- 2021-08-09 WO PCT/CN2021/111511 patent/WO2023000403A1/zh unknown
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107708671A (zh) * | 2015-04-16 | 2018-02-16 | 纽约州立大学研究基金会 | 包含钴卟啉‑磷脂缀合物和聚组氨酸标签的纳米结构 |
CN106924731A (zh) * | 2015-12-30 | 2017-07-07 | 复旦大学 | 一种基于光动力疗法和化学疗法的联合肿瘤靶向治疗系统 |
CN109464395A (zh) * | 2019-01-03 | 2019-03-15 | 中国科学院过程工程研究所 | 一种水包油包凝胶乳液及其制备方法和应用 |
CN112569207A (zh) * | 2019-09-30 | 2021-03-30 | 复旦大学 | 一种载脂蛋白修饰的仿生纳米肿瘤疫苗及其制备方法和用途 |
CN111217917A (zh) * | 2020-02-26 | 2020-06-02 | 康希诺生物股份公司 | 一种新型冠状病毒SARS-CoV-2疫苗及其制备方法 |
CN111560074A (zh) * | 2020-03-20 | 2020-08-21 | 中山大学 | 一种基于幽门螺旋杆菌铁蛋白的新型冠状病毒s蛋白单区域亚单位纳米疫苗 |
Non-Patent Citations (1)
Title |
---|
WEI-CHIAO HUANG等: "SARS-CoV-2 RBD Neutralizing Antibody Induction is Enhanced by Particulate Vaccination", 《ADV MATER》 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023000403A1 (zh) * | 2021-07-20 | 2023-01-26 | 武汉圣润生物科技有限公司 | 一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用 |
CN114344457A (zh) * | 2021-11-13 | 2022-04-15 | 暨南大学 | 新型冠状病毒蛋白质抗原纳米疫苗及其制备方法和应用 |
CN114344457B (zh) * | 2021-11-13 | 2024-01-12 | 暨南大学 | 新型冠状病毒蛋白质抗原纳米疫苗及其制备方法和应用 |
CN115969969A (zh) * | 2023-02-13 | 2023-04-18 | 中国科学院长春应用化学研究所 | 一种仿病毒结构纳米颗粒疫苗及其制备方法和应用 |
Also Published As
Publication number | Publication date |
---|---|
WO2023000403A1 (zh) | 2023-01-26 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Li et al. | Rational design of polymeric hybrid micelles to overcome lymphatic and intracellular delivery barriers in cancer immunotherapy | |
Zhang et al. | Peptide amphiphile micelle vaccine size and charge influence the host antibody response | |
US11510996B2 (en) | Covalent polymer-antigen conjugated particles | |
US20080160089A1 (en) | Vaccine delivery compositions and methods of use | |
US8017154B2 (en) | Polyamino acid for use as adjuvant | |
CN113476598A (zh) | 一种新型冠状病毒亚蛋白纳米疫苗及其制备方法和应用 | |
CN102202653B (zh) | 用于免疫疗法的纳米颗粒 | |
Moura et al. | Functionalized branched polymers: promising immunomodulatory tools for the treatment of cancer and immune disorders | |
WO2007089870A2 (en) | Vaccine delivery compositions and methods of use | |
US20070160622A1 (en) | Method for assembling a polymer-biologic delivery composition | |
Yan et al. | An overview of biodegradable nanomaterials and applications in vaccines | |
TW200307557A (en) | Stabilized synthetic immunogen delivery systems | |
KR20070101341A (ko) | 백신 전달 조성물 및 사용 방법 | |
US20060188469A1 (en) | Vaccine delivery compositions and methods of use | |
EP3210621B1 (en) | Micellar polypeptide vaccine having pegylated phospholipids as carrier | |
Ebrahimian et al. | Induction of a balanced Th1/Th2 immune responses by co-delivery of PLGA/ovalbumin nanospheres and CpG ODNs/PEI-SWCNT nanoparticles as TLR9 agonist in BALB/c mice | |
Yang et al. | Synthetic, supramolecular, and self‐adjuvanting CD8+ T‐cell epitope vaccine increases the therapeutic antitumor immunity | |
Liu et al. | Antigen-conjugated N-trimethylaminoethylmethacrylate chitosan nanoparticles induce strong immune responses after nasal administration | |
Song et al. | Photoresponsive polypeptide-glycosylated dendron amphiphiles: UV-triggered polymersomes, OVA release, and in vitro enhanced uptake and immune response | |
Shen et al. | Nano-vesicles based on phospholipid-like amphiphilic polyphosphazenes to orally deliver ovalbumin antigen for evoking anti-tumor immune response | |
Lin et al. | Enhanced immune responses to mucosa by functionalized chitosan-based composite nanoparticles as a vaccine adjuvant for intranasal delivery | |
Malek-Khatabi et al. | Long-term vaccine delivery and immunological responses using biodegradable polymer-based carriers | |
CN101506266A (zh) | 疫苗输送组合物及使用方法 | |
Li et al. | Emerging nanoparticle platforms for CpG oligonucleotide delivery | |
WO2006112476A2 (ja) | 抗原を固定化した生分解性ナノ粒子、およびそれを含むワクチン |
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 |