CN104918637A - 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 - Google Patents
用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 Download PDFInfo
- Publication number
- CN104918637A CN104918637A CN201380063896.1A CN201380063896A CN104918637A CN 104918637 A CN104918637 A CN 104918637A CN 201380063896 A CN201380063896 A CN 201380063896A CN 104918637 A CN104918637 A CN 104918637A
- Authority
- CN
- China
- Prior art keywords
- seq
- fusion rotein
- cell
- antigen
- aminoacid sequence
- 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
- 239000000427 antigen Substances 0.000 title claims abstract description 78
- 108091007433 antigens Proteins 0.000 title claims abstract description 78
- 102000036639 antigens Human genes 0.000 title claims abstract description 78
- 230000001939 inductive effect Effects 0.000 title claims description 5
- 230000005867 T cell response Effects 0.000 title abstract description 7
- 108020001507 fusion proteins Proteins 0.000 title abstract description 6
- 102000037865 fusion proteins Human genes 0.000 title abstract description 6
- 230000002163 immunogen Effects 0.000 title abstract description 3
- 239000003623 enhancer Substances 0.000 title abstract 2
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 108
- 230000004927 fusion Effects 0.000 claims abstract description 93
- 210000001744 T-lymphocyte Anatomy 0.000 claims abstract description 79
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 63
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 49
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 47
- 210000000612 antigen-presenting cell Anatomy 0.000 claims abstract description 39
- 230000026683 transduction Effects 0.000 claims abstract description 32
- 238000010361 transduction Methods 0.000 claims abstract description 32
- 244000052769 pathogen Species 0.000 claims abstract description 29
- 230000001717 pathogenic effect Effects 0.000 claims abstract description 29
- 229920001184 polypeptide Polymers 0.000 claims abstract description 27
- 101001043564 Homo sapiens Prolow-density lipoprotein receptor-related protein 1 Proteins 0.000 claims abstract description 21
- 125000000539 amino acid group Chemical group 0.000 claims abstract description 16
- 230000002708 enhancing effect Effects 0.000 claims abstract description 7
- 108700033844 Pseudomonas aeruginosa toxA Proteins 0.000 claims description 57
- 206010070834 Sensitisation Diseases 0.000 claims description 44
- 230000008313 sensitization Effects 0.000 claims description 44
- 235000018102 proteins Nutrition 0.000 claims description 42
- 229960005486 vaccine Drugs 0.000 claims description 42
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 40
- 210000004027 cell Anatomy 0.000 claims description 39
- 230000019491 signal transduction Effects 0.000 claims description 33
- 210000002472 endoplasmic reticulum Anatomy 0.000 claims description 22
- 230000014759 maintenance of location Effects 0.000 claims description 18
- 108700018351 Major Histocompatibility Complex Proteins 0.000 claims description 15
- 230000020382 suppression by virus of host antigen processing and presentation of peptide antigen via MHC class I Effects 0.000 claims description 15
- 230000000890 antigenic effect Effects 0.000 claims description 14
- 201000010099 disease Diseases 0.000 claims description 13
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 13
- 239000002671 adjuvant Substances 0.000 claims description 11
- 210000000170 cell membrane Anatomy 0.000 claims description 6
- 208000015181 infectious disease Diseases 0.000 claims description 6
- 208000024891 symptom Diseases 0.000 claims description 6
- JBFQOLHAGBKPTP-NZATWWQASA-N (2s)-2-[[(2s)-4-carboxy-2-[[3-carboxy-2-[[(2s)-2,6-diaminohexanoyl]amino]propanoyl]amino]butanoyl]amino]-4-methylpentanoic acid Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)C(CC(O)=O)NC(=O)[C@@H](N)CCCCN JBFQOLHAGBKPTP-NZATWWQASA-N 0.000 claims description 3
- 108010089256 lysyl-aspartyl-glutamyl-leucine Proteins 0.000 claims description 3
- 230000005945 translocation Effects 0.000 abstract 2
- 230000001235 sensitizing effect Effects 0.000 abstract 1
- 102100036011 T-cell surface glycoprotein CD4 Human genes 0.000 description 19
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 description 17
- 241000699670 Mus sp. Species 0.000 description 16
- 241000701806 Human papillomavirus Species 0.000 description 13
- 206010028980 Neoplasm Diseases 0.000 description 12
- 241001135989 Porcine reproductive and respiratory syndrome virus Species 0.000 description 12
- 239000003795 chemical substances by application Substances 0.000 description 11
- 241001465754 Metazoa Species 0.000 description 10
- 229960001438 immunostimulant agent Drugs 0.000 description 10
- 239000003022 immunostimulating agent Substances 0.000 description 10
- 230000003308 immunostimulating effect Effects 0.000 description 10
- 238000000034 method Methods 0.000 description 10
- 210000004988 splenocyte Anatomy 0.000 description 10
- NKVLDFAVEWLOCX-GUSKIFEASA-N [(2s,3r,4s,5r,6r)-3-[(2s,3r,4s,5r,6s)-5-[(2s,3r,4s,5r)-4-[(2s,3r,4r)-3,4-dihydroxy-4-(hydroxymethyl)oxolan-2-yl]oxy-3,5-dihydroxyoxan-2-yl]oxy-3,4-dihydroxy-6-methyloxan-2-yl]oxy-4,5-dihydroxy-6-methyloxan-2-yl] (4ar,5r,6as,6br,9s,10s,12ar)-10-[(2r,3r,4s, Chemical compound O([C@H]1[C@H](O)CO[C@H]([C@@H]1O)O[C@H]1[C@H](C)O[C@H]([C@@H]([C@@H]1O)O)O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](C)O[C@H]1OC(=O)[C@]12CCC(C)(C)CC1C1=CCC3[C@@]([C@@]1(C[C@H]2O)C)(C)CCC1[C@]3(C)CC[C@@H]([C@@]1(C)C=O)O[C@@H]1O[C@@H]([C@H]([C@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)CO2)O)[C@H]1O[C@H]1[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O1)O)O)C(=O)NCCCCCCCCCCCC)[C@@H]1OC[C@](O)(CO)[C@H]1O NKVLDFAVEWLOCX-GUSKIFEASA-N 0.000 description 9
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
- 239000012634 fragment Substances 0.000 description 9
- 230000005847 immunogenicity Effects 0.000 description 9
- 238000011081 inoculation Methods 0.000 description 9
- 239000000902 placebo Substances 0.000 description 9
- 229940068196 placebo Drugs 0.000 description 9
- ILRRQNADMUWWFW-UHFFFAOYSA-K aluminium phosphate Chemical compound O1[Al]2OP1(=O)O2 ILRRQNADMUWWFW-UHFFFAOYSA-K 0.000 description 8
- 239000013604 expression vector Substances 0.000 description 8
- 239000013612 plasmid Substances 0.000 description 8
- 108020003175 receptors Proteins 0.000 description 8
- 102000005962 receptors Human genes 0.000 description 8
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 8
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 description 7
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 229940123205 CD28 agonist Drugs 0.000 description 6
- 241001673669 Porcine circovirus 2 Species 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 210000004443 dendritic cell Anatomy 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 6
- 230000005760 tumorsuppression Effects 0.000 description 6
- 102000004172 Cathepsin L Human genes 0.000 description 5
- 108090000624 Cathepsin L Proteins 0.000 description 5
- 108010074328 Interferon-gamma Proteins 0.000 description 5
- 238000003776 cleavage reaction Methods 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 230000003053 immunization Effects 0.000 description 5
- 230000006698 induction Effects 0.000 description 5
- 230000001404 mediated effect Effects 0.000 description 5
- 230000007017 scission Effects 0.000 description 5
- 238000001179 sorption measurement Methods 0.000 description 5
- 238000010254 subcutaneous injection Methods 0.000 description 5
- 239000007929 subcutaneous injection Substances 0.000 description 5
- 108090001126 Furin Proteins 0.000 description 4
- 102000004961 Furin Human genes 0.000 description 4
- 241000711549 Hepacivirus C Species 0.000 description 4
- 108700039791 Hepatitis C virus nucleocapsid Proteins 0.000 description 4
- 102000035195 Peptidases Human genes 0.000 description 4
- 108091005804 Peptidases Proteins 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 230000021615 conjugation Effects 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 108700025184 hepatitis B virus X Proteins 0.000 description 4
- 238000002649 immunization Methods 0.000 description 4
- 230000000638 stimulation Effects 0.000 description 4
- 230000017105 transposition Effects 0.000 description 4
- 102100027165 Alpha-2-macroglobulin receptor-associated protein Human genes 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 241000700721 Hepatitis B virus Species 0.000 description 3
- 241000725303 Human immunodeficiency virus Species 0.000 description 3
- 241000341655 Human papillomavirus type 16 Species 0.000 description 3
- 102000008070 Interferon-gamma Human genes 0.000 description 3
- 102100021923 Prolow-density lipoprotein receptor-related protein 1 Human genes 0.000 description 3
- 238000013459 approach Methods 0.000 description 3
- 210000003719 b-lymphocyte Anatomy 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 3
- 238000013467 fragmentation Methods 0.000 description 3
- 238000006062 fragmentation reaction Methods 0.000 description 3
- 230000000799 fusogenic effect Effects 0.000 description 3
- 229940044627 gamma-interferon Drugs 0.000 description 3
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 3
- 230000010534 mechanism of action Effects 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000005728 strengthening Methods 0.000 description 3
- 102100033312 Alpha-2-macroglobulin Human genes 0.000 description 2
- 108091035707 Consensus sequence Proteins 0.000 description 2
- 241000725619 Dengue virus Species 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 102000002812 Heat-Shock Proteins Human genes 0.000 description 2
- 108010004889 Heat-Shock Proteins Proteins 0.000 description 2
- 108010070875 Human Immunodeficiency Virus tat Gene Products Proteins 0.000 description 2
- 102100037850 Interferon gamma Human genes 0.000 description 2
- NPBGTPKLVJEOBE-IUCAKERBSA-N Lys-Arg Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(O)=O)CCCNC(N)=N NPBGTPKLVJEOBE-IUCAKERBSA-N 0.000 description 2
- 102000005431 Molecular Chaperones Human genes 0.000 description 2
- 108010006519 Molecular Chaperones Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 101000596353 Severe acute respiratory syndrome coronavirus 2 ORF7a protein Proteins 0.000 description 2
- 210000000447 Th1 cell Anatomy 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- 239000000556 agonist Substances 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 235000013877 carbamide Nutrition 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 230000020411 cell activation Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 2
- 230000012202 endocytosis Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000008105 immune reaction Effects 0.000 description 2
- 210000003000 inclusion body Anatomy 0.000 description 2
- 229930027917 kanamycin Natural products 0.000 description 2
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 2
- 229960000318 kanamycin Drugs 0.000 description 2
- 229930182823 kanamycin A Natural products 0.000 description 2
- 210000003712 lysosome Anatomy 0.000 description 2
- 230000001868 lysosomic effect Effects 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 210000005087 mononuclear cell Anatomy 0.000 description 2
- 230000026938 proteasomal ubiquitin-dependent protein catabolic process Effects 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000002864 sequence alignment Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 230000004614 tumor growth Effects 0.000 description 2
- 241000712461 unidentified influenza virus Species 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- 101000621943 Acholeplasma phage L2 Probable integrase/recombinase Proteins 0.000 description 1
- 101000618348 Allochromatium vinosum (strain ATCC 17899 / DSM 180 / NBRC 103801 / NCIMB 10441 / D) Uncharacterized protein Alvin_0065 Proteins 0.000 description 1
- 101000636214 Arabidopsis thaliana Transcription factor MYC2 Proteins 0.000 description 1
- 101000781117 Autographa californica nuclear polyhedrosis virus Uncharacterized 12.4 kDa protein in CTL-LEF2 intergenic region Proteins 0.000 description 1
- 101000708323 Azospirillum brasilense Uncharacterized 28.8 kDa protein in nifR3-like 5'region Proteins 0.000 description 1
- 101000770311 Azotobacter chroococcum mcd 1 Uncharacterized 19.8 kDa protein in nifW 5'region Proteins 0.000 description 1
- 101000748761 Bacillus subtilis (strain 168) Uncharacterized MFS-type transporter YcxA Proteins 0.000 description 1
- 101000765620 Bacillus subtilis (strain 168) Uncharacterized protein YlxP Proteins 0.000 description 1
- 101000916134 Bacillus subtilis (strain 168) Uncharacterized protein YqxJ Proteins 0.000 description 1
- 241000255789 Bombyx mori Species 0.000 description 1
- 101000754349 Bordetella pertussis (strain Tohama I / ATCC BAA-589 / NCTC 13251) UPF0065 protein BP0148 Proteins 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 101000827633 Caldicellulosiruptor sp. (strain Rt8B.4) Uncharacterized 23.9 kDa protein in xynA 3'region Proteins 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 101000947628 Claviceps purpurea Uncharacterized 11.8 kDa protein Proteins 0.000 description 1
- 101000686796 Clostridium perfringens Replication protein Proteins 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 101150013359 E7 gene Proteins 0.000 description 1
- 101000788129 Escherichia coli Uncharacterized protein in sul1 3'region Proteins 0.000 description 1
- 101000788370 Escherichia phage P2 Uncharacterized 12.9 kDa protein in GpA 3'region Proteins 0.000 description 1
- 101000787096 Geobacillus stearothermophilus Uncharacterized protein in gldA 3'region Proteins 0.000 description 1
- 101000976889 Haemophilus phage HP1 (strain HP1c1) Uncharacterized 19.2 kDa protein in cox-rep intergenic region Proteins 0.000 description 1
- 101000767631 Human papillomavirus type 16 Protein E7 Proteins 0.000 description 1
- 101000827627 Klebsiella pneumoniae Putative low molecular weight protein-tyrosine-phosphatase Proteins 0.000 description 1
- 108010003718 LDL-Receptor Related Protein-Associated Protein Proteins 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 101001130841 Middle East respiratory syndrome-related coronavirus (isolate United Kingdom/H123990006/2012) Non-structural protein ORF5 Proteins 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 101710087110 ORF6 protein Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 108010004729 Phycoerythrin Proteins 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 101000974028 Rhizobium leguminosarum bv. viciae (strain 3841) Putative cystathionine beta-lyase Proteins 0.000 description 1
- 101000756519 Rhodobacter capsulatus (strain ATCC BAA-309 / NBRC 16581 / SB1003) Uncharacterized protein RCAP_rcc00048 Proteins 0.000 description 1
- 101000948219 Rhodococcus erythropolis Uncharacterized 11.5 kDa protein in thcD 3'region Proteins 0.000 description 1
- 101000936711 Streptococcus gordonii Accessory secretory protein Asp4 Proteins 0.000 description 1
- 101000929863 Streptomyces cinnamonensis Monensin polyketide synthase putative ketoacyl reductase Proteins 0.000 description 1
- 101000788468 Streptomyces coelicolor Uncharacterized protein in mprR 3'region Proteins 0.000 description 1
- 101000845085 Streptomyces violaceoruber Granaticin polyketide synthase putative ketoacyl reductase 1 Proteins 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 235000009470 Theobroma cacao Nutrition 0.000 description 1
- 101000711771 Thiocystis violacea Uncharacterized 76.5 kDa protein in phbC 3'region Proteins 0.000 description 1
- 101710095001 Uncharacterized protein in nifU 5'region Proteins 0.000 description 1
- 101000711318 Vibrio alginolyticus Uncharacterized 11.6 kDa protein in scrR 3'region Proteins 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 210000005006 adaptive immune system Anatomy 0.000 description 1
- 230000030741 antigen processing and presentation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 201000008275 breast carcinoma Diseases 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000008366 buffered solution Substances 0.000 description 1
- 244000240602 cacao Species 0.000 description 1
- 239000002458 cell surface marker Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000021633 leukocyte mediated immunity Effects 0.000 description 1
- 108010045758 lysosomal proteins Proteins 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 230000010837 receptor-mediated endocytosis Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000012744 reinforcing agent Substances 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
- 229940031626 subunit vaccine Drugs 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 230000001550 time effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
-
- 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
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/38—Albumins
- A61K38/385—Serum albumin
-
- 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
- A61K39/125—Picornaviridae, e.g. calicivirus
- A61K39/135—Foot- and mouth-disease virus
-
- 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
- A61K39/155—Paramyxoviridae, e.g. parainfluenza virus
- A61K39/17—Newcastle disease virus
-
- 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
- A61K39/21—Retroviridae, e.g. equine infectious anemia virus
-
- 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
- A61K39/29—Hepatitis virus
-
- 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
- A61K39/29—Hepatitis virus
- A61K39/292—Serum hepatitis virus, hepatitis B virus, e.g. Australia 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
- A61K39/385—Haptens or antigens, bound to carriers
-
- 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
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
- A61K47/646—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- 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
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- 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
-
- 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
- A61P31/04—Antibacterial agents
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
-
- 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
- A61P31/18—Antivirals for RNA viruses for HIV
-
- 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
- A61P31/20—Antivirals for DNA viruses
-
- 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
- A61P31/20—Antivirals for DNA viruses
- A61P31/22—Antivirals for DNA viruses for herpes viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70596—Molecules with a "CD"-designation not provided for elsewhere
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/81—Protease inhibitors
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/82—Translation products from oncogenes
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1048—Glycosyltransferases (2.4)
- C12N9/1077—Pentosyltransferases (2.4.2)
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y204/00—Glycosyltransferases (2.4)
- C12Y204/02—Pentosyltransferases (2.4.2)
- C12Y204/02036—NAD(+)--diphthamide ADP-ribosyltransferase (2.4.2.36)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01018—Exo-alpha-sialidase (3.2.1.18), i.e. trans-sialidase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y306/00—Hydrolases acting on acid anhydrides (3.6)
- C12Y306/05—Hydrolases acting on acid anhydrides (3.6) acting on GTP; involved in cellular and subcellular movement (3.6.5)
- C12Y306/05002—Small monomeric GTPase (3.6.5.2)
-
- 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
-
- 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
- A61K2039/58—Medicinal preparations containing antigens or antibodies raising an immune response against a target which is not the antigen used for immunisation
- A61K2039/585—Medicinal preparations containing antigens or antibodies raising an immune response against a target which is not the antigen used for immunisation wherein the target is cancer
-
- 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
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
- A61K2039/6031—Proteins
- A61K2039/6037—Bacterial toxins, e.g. diphteria toxoid [DT], tetanus toxoid [TT]
-
- 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
- A61K2039/6043—Heat shock proteins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/21—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Pseudomonadaceae (F)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/04—Fusion polypeptide containing a localisation/targetting motif containing an ER retention signal such as a C-terminal HDEL motif
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/095—Fusion polypeptide containing a localisation/targetting motif containing a nuclear export signal
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/33—Fusion polypeptide fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/55—Fusion polypeptide containing a fusion with a toxin, e.g. diphteria toxin
-
- 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
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/12011—Asfarviridae
- C12N2710/12022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- 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
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/20011—Papillomaviridae
- C12N2710/20022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- 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
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/20011—Papillomaviridae
- C12N2710/20034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- 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
- C12N2730/00—Reverse transcribing DNA viruses
- C12N2730/00011—Details
- C12N2730/10011—Hepadnaviridae
- C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
- C12N2730/10122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- 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
- C12N2730/00—Reverse transcribing DNA viruses
- C12N2730/00011—Details
- C12N2730/10011—Hepadnaviridae
- C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
- C12N2730/10134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- 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
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/16011—Human Immunodeficiency Virus, HIV
- C12N2740/16022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- 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
- C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
- C12N2750/00011—Details
- C12N2750/10011—Circoviridae
- C12N2750/10022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- 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
- C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
- C12N2750/00011—Details
- C12N2750/10011—Circoviridae
- C12N2750/10034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/18011—Paramyxoviridae
- C12N2760/18111—Avulavirus, e.g. Newcastle disease virus
- C12N2760/18122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/18011—Paramyxoviridae
- C12N2760/18111—Avulavirus, e.g. Newcastle disease virus
- C12N2760/18134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- 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
- C12N2770/24011—Flaviviridae
- C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
- C12N2770/24222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- 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
- C12N2770/24011—Flaviviridae
- C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
- C12N2770/24234—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- 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
- C12N2770/24011—Flaviviridae
- C12N2770/24311—Pestivirus, e.g. bovine viral diarrhea virus
- C12N2770/24334—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- 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
- C12N2770/32011—Picornaviridae
- C12N2770/32111—Aphthovirus, e.g. footandmouth disease virus
- C12N2770/32122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- 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
- C12N2770/32011—Picornaviridae
- C12N2770/32111—Aphthovirus, e.g. footandmouth disease virus
- C12N2770/32134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
本申请公开一种用作增强抗原特异性T细胞反应的免疫原增强剂的融合蛋白。所述融合蛋白包含:(a)抗原呈递细胞(APC)结合域或CD91受体结合域;(b)蛋白转导域;及(c)病原体的抗原,其中,所述APC结合域或CD91受体结合域位于所述融合蛋白的N端,且所述病原体的抗原位于所述蛋白转导域的C端。所述蛋白转导域选自:(i)融合多肽,其包含T细胞致敏信号转导肽、连接子及转位肽;(it)T细胞致敏信号转导肽;及(iii)长度为34-112个氨基酸残基的转位肽。
Description
技术领域
本发明主要涉及融合蛋白,并且更具体地涉及增强T细胞介导的免疫反应的融合蛋白。
背景技术
分子生物学已使亚单位疫苗的生产成为可能,其中,免疫原为母蛋白或母复合物的片段或亚单位。所需要的是可诱发T细胞致敏反应又能足够灵活的并入来自许多感染源的毒株的序列的稳定疫苗的开发。
发明内容
一方面,本发明涉及一种融合蛋白,其包含:
(a)抗原呈递细胞(APC)结合域或CD91受体结合域,其位于所述融合蛋白的N端;
(b)蛋白转导域,其位于所述APC结合域或CD91受体结合域的C端,所述蛋白转导域选自:
(i)融合多肽,其包含T细胞致敏信号转导肽、连接子及转位肽,其中:
(1)所述T细胞致敏信号转导肽位于所述融合多肽的N端;
(2)所述连接子包含SEQ ID NO:15,其连接所述T细胞致敏信号转导胜肽与所述转位肽;且
(3)所述转位肽的长度为34-112个氨基酸残基,且包含与SEQID NO:3、SEQ ID NO:20或SEQ ID NO:4至少90%相同的氨基酸序列;
(ii)T细胞致敏信号转导肽;及
(iii)长度为34-112个氨基酸残基的转位肽,其包含与SEQ ID NO:3、SEQ ID NO:20或SEQ ID NO:4至少90%相同的氨基酸序列;及
(c)病原体的抗原,其位于所述蛋白转导域的C端;
其中:
所述T细胞致敏信号转导肽的长度为28-53个氨基酸残基,且包含与SEQID NO:31至少90%相同的氨基酸序列,其中Xaa8为I或L;Xaa10为V、F或A;Xaa11为M或L;Xaa17为L或I;且
若所述蛋白转导域为所述转位肽(biii),则所述APC结合域或CD91受体结合域无假单胞菌外毒素A(PE)结合域I的氨基酸序列。
在本发明的一个实施方式中,所述APC结合域或所述CD91受体结合域为包含与选自SEQ ID NO:5、SEQ ID NO:9、SEQ ID NO:6、SEQ ID NO:7及SEQ ID NO:8的序列至少90%相同的氨基酸序列的多肽。或者,所述APC结合域选自受体相关蛋白-1(RAP1)区域III、α-2-巨球蛋白受体相关蛋白(A2M)、HIV-Tat、热休克蛋白(HSP)及假单胞菌外毒素A(PE)结合域I。
在本发明的另一个实施方式中,所述融合蛋白无假单胞菌外毒素A(PE)结合域I的氨基酸序列。
在本发明的另一个实施方式中,所述融合蛋白进一步包含位于所述融合蛋白C端的内质网(ER)滞留序列。
在本发明的另一个实施方式中,所述内质网滞留序列包含氨基酸序列Lys-Asp-Glu-Leu(SEQ ID NO:14)。所述ER滞留序列可包含选自SEQ ID NO:14、SEQ ID NO:16至SEQ ID NO:19的序列。或者,所述ER滞留序列可由选自SEQ ID NO:16至SEQ ID NO:19的序列组成。
在本发明的另一个实施方式中,若所述抗原含有10个或更多抗原决定簇,则所述融合蛋白的C端无内质网滞留序列。
在本发明的另一个实施方式中,所述蛋白转导域为融合多肽(bi)。
在本发明的另一个实施方式中,所述蛋白转导域为T细胞致敏信号转导肽(bii)。
在本发明的另一个实施方式中,所述融合蛋白在所述蛋白转导域与所述抗原之间进一步包含另一连接子,所述另一连接子包含SEQ ID NO:15。
在本发明的另一个实施方式中,所述蛋白转导域为转位肽(biii)。
在本发明的另一个实施例方式中,所述融合蛋白在所述APC结合域或所述CD91受体结合域与所述转位肽之间进一步包含另一连接子,所述另一连接子包含SEQ ID NO:15。
在本发明的另一个实施方式中,所述蛋白转导域包含SEQ ID NO:30。
在本发明的另一个实施方式中,所述APC结合域包含与选自SEQ ID NO:5、SEQ ID NO:9、SEQ ID NO:6、SEQ ID NO:7及SEQ ID NO:8的序列至少95%相同的氨基酸序列。
在本发明的另一个实施方式中,所述APC结合域或所述CD91受体结合域为包含选自SEQ ID NO:5、SEQ ID NO:9、SEQ ID NO:6、SEQ ID NO:7及SEQ ID NO:8的氨基酸序列的多肽。
在本发明的另一个实施方式中,所述T细胞致敏信号转导肽包含与SEQID NO:1或SEQ ID NO:2至少90%相同的氨基酸序列。
在本发明的另一个实施方式中,所述T细胞致敏信号转导肽包含选自SEQ ID NO:1及SEQ ID NO:2的氨基酸序列。
在本发明的另一个实施方式中,所述转位肽包含选自SEQ ID NO:3、SEQID NO:20及SEQ ID NO:4的氨基酸序列。
在本发明的另一个实施方式中,所述转位肽的长度为34-61个氨基酸残基。
在本发明的另一个实施方式中,前述融合蛋白的蛋白转导域具有以下特征:(i)所述T细胞致敏信号转导肽包含SEQ ID NO:1或SEQ ID NO:2的氨基酸序列;且(ii)所述转位肽包含与SEQ ID NO:3至少95%相同的氨基酸序列。
所述T细胞致敏信号转导肽显示出诱发识别并结合于T细胞上CD28受体的K1(X)2E3(X)4(X)5Y6P7P8P9Y10氨基酸序列(SEQ ID NO:32)的抗体的特征,其中,(X)2为I或L;(X)4为V、F或A;且(X)5为M或L。
另一方面,本发明涉及一种融合蛋白,其由下列各项所组成:
(a)抗原呈递细胞(APC)结合域或CD91受体结合域,其位于所述融合蛋白的N端;
(b)蛋白转导域,其位于所述APC结合域或CD91受体结合域的C端,所述蛋白转导域选自:
(i)融合多肽,其包含T细胞致敏信号转导肽、连接子及转位肽,其中:
(1)所述T细胞致敏信号转导肽位于所述融合多肽的N端;
(2)所述连接子包含SEQ ID NO:15,其连接所述T细胞致敏信号转导肽与所述转位肽;且
(3)所述转位肽的长度为34-112个氨基酸残基,其包含与SEQID NO:3、SEQ ID NO:20或SEQ ID NO:4至少90%相同的氨基酸序列;
(ii)T细胞致敏信号转导肽;及
(iii)34-112个氨基酸残基长度的转位肽,,其包含与SEQ ID NO:3、20或4号至少90%相同的氨基酸序列;及
(c)病原体的抗原,其位于所述蛋白转导域的C端;
其中:
所述T细胞致敏信号转导肽的长度为28-53个氨基酸残基,且包含与SEQID NO:31至少90%相同的氨基酸序列,其中,Xaa8为I或L;Xaa10为V、F或A;Xaa11为M或L;Xaa17为L或I;且
若所述蛋白转导域为所述转位肽(biii),则所述APC结合域或所述CD91受体结合域无假单胞菌外毒素A(PE)结合域I的氨基酸序列。
所述抗原呈递细胞(APC)可以选自树突细胞、巨噬细胞、B细胞及单核细胞。
在本发明的一个实施方式中,所述APC的细胞膜包含CD91受体。
另一方面,本发明涉及一种疫苗组合物,其包含:(a)治疗有效量的前述融合蛋白;及(b)佐剂。
所述佐剂为抗原传送剂或免疫增效剂。在本发明的一个实施方式中,所述疫苗组合物包含抗原传送剂而无免疫增效剂。
此外,在另一方面,本发明涉及一种诱导增强的病原体抗原特异性T细胞反应的方法,其包括:对有此需要的受试者施用包含治疗有效量的前述融合蛋白的疫苗组合物;以及由此诱导增强的病原体抗原特异性T细胞反应。
此外,在另一方面,本发明涉及一种杀死通过疾病细胞的细胞膜上的第I类主要组织相容性复合体(MHC)分子呈递抗原的疾病细胞的方法,其包括:对有此需要的受试者施用疫苗组合物,所述疫苗组合物包含治疗有效量的前述融合蛋白;以及由此杀死通过疾病细胞的细胞膜上的第I类MHC分子呈递抗原的疾病细胞。
在本发明的一个实施方式中,所述疾病细胞为癌细胞。
在又一个方面,本发明涉及一种预防、治疗由病原体所引起的感染和/或将由感染所引起的症状减至最少的方法,其包括:对有此需要的受试者施用疫苗组合物,所述疫苗组合物包含治疗有效量的权利要求1的融合蛋白;以及由此预防、治疗由病原体所引起的感染和/或将由感染所引起的症状减至最少。本发明还涉及前述融合蛋白或疫苗组合物,其用于诱导增强的病原体抗原特异性T细胞反应,或用于杀死通过疾病细胞的细胞膜上的第I类MHC分子呈递抗原的疾病细胞,或用于预防、治疗由病原体所引起的感染和/或将由感染所引起的症状减至最少。
所述病原体可以是选自人乳头瘤病毒(HPV)、猪繁殖与呼吸综合症病毒(PRRSV)、人免疫缺陷病毒(HIV-1)、流感病毒、登革热病毒、丙型肝炎病毒(HCV)、乙型肝炎病毒(HBV)及猪环状病毒2(PCV2)中的至少一种。
在本发明的一个实施方式中,前述的融合蛋白为用于增强有此需要的受试者的抗原特异性细胞毒性T细胞反应。所述融合蛋白在有此需要的受试者体内,亦可用于增强抗原特异性CD4+T细胞反应,或用作诱导增强的抗原特异性抗体效价反应的免疫原性增效剂。
由下述结合以下附图的优选实施方式的描述将明显可见这些及其他方面,但是在不偏离本说明书的新概念的实质和范围的情况下可影响其中的变化和修改。
附图显示本发明的一种或多种实施方式,并与书面说明共同用于解释本发明的原理。在可能情况下,整个附图使用的附图标记指实施例的相同或相似要素。
附图说明
图1为载体图谱。
图2为显示融合蛋白的SDS-PAGE分析结果的照片。
图3为载体图谱。
图4为显示本发明的一个实施方式的示意图。
图5A显示免疫接种程序表。
图5B为显示融合蛋白的SDS-PAGE分析结果的照片。
图5C-D分别为显示接种不同融合蛋白或安慰剂的动物组的肿瘤大小曲线及无肿瘤小鼠的百分比。
图6为制备融合蛋白的流程图(左图),以及显示融合蛋白的SDS-PAGE分析结果的照片(右图)。
图7为显示T细胞致敏融合蛋白作用机理的示意图。
图8显示不同物种的CD28的序列比对。
图9A显示免疫接种程序表。
图9B-C分别显示接种不同融合蛋白或安慰剂的动物组的肿瘤大小曲线及存活率。
图10为显示用于由病原体产生含有DNA插入片段的质粒的含有RAP1的载体的示意图。
图11为显示含有各种病原体的抗原的融合蛋白结构的示意图。
图12A-F为显示各种融合蛋白的SDS-PAGE分析结果的照片。
图13A-B显示动物组、用于免疫动物的疫苗与剂量,以及免疫接种程序表。
图14A-D为显示来自已接种安慰剂或含有E716、E718、HCV核心或HBx抗原的融合蛋白的图13A的动物组的CD3+/CD4+脾细胞及CD3+/CD8+脾细胞的体外(ex vivo)抗原特异性免疫反应的分析结果的表格。
图15A-J显示来自已接种安慰剂或含有PCV2(图15A-B)或PRRSV抗原(图15C-J)的融合蛋白的图13A的动物组的CD3+/CD8+脾细胞及CD3+/CD4+脾细胞的体外抗原特异性免疫反应分析的IFNγ+细胞计数。
具体实施方式
在以下仅意欲作为说明的实施例中更具体地描述本发明,因为对本领域的技术人员来说其中的修改和变化将明显可见。现在,详细描述本发明的各种实施方式。参考附图,整个图中相同标号表示相同部分。当在此用于说明书和接着的整个权利要求时,除非上下文另外载明,否则“一”、“一个”及“该”的意思包括多个引用项目。并且,当在此用于说明书和接着的整个权利要求时,除非上下文另外载明,否则“在…中”的意思包括“在…中”及「“在…上”。此外,为读者的便利在说明书中可以使用标题及小标题,其不应当影响本发明的范围。因此,以下将更具体地定义用于本说明书的若干术语。
定义
用于本说明书的术语在本发明的上下文中及使用各术语的特定上下文中通常在本领域中具有它们的通常涵义。以下或在本说明书的其他地方将讨论用以描述本发明的某些术语以向从业者提供关于本发明的描述的额外指导。为便利,某些术语可能例如使用斜体和/或引号来突出显示。突出显示的使用不影响术语的范围与意义;在相同上下文中术语的范围与意义相同,无论是否突出显示。将意识到同一件事能够以不止一种方式表达。因此,可选择的语言和同义词可以用于在此讨论的任一一种或多种术语,并无任何特殊意义强加于是否在此详细说明或讨论术语。提供某些术语的同义词。一种或多种同义词的列举不排除其他同义词的使用。本说明书中任何地方的实施例的使用(包括在此所讨论的术语的实施例)仅供说明之用,而非用于限制本发明或任何示例性术语的范围与意义。同样地,本发明不局限于在本说明书中给出的各种实施方式。
除非另有定义,在此使用的所有技术与科学术语与本领域中关于本发明的普通技术人员中的一个通常所理解的具有相同含义。若有相互冲突之处,则以本说明书及其所提供的定义为解释依据。
如在此使用的,“左右”、“约”或“大约”通常指在给出的数值或范围的20%以内,优选在10%以内,并且更优选在5%以内。在此给出的数量是大约的,指如果没明确规定能够推测术语“左右”、“约”或“大约”。
术语“抗原呈递细胞(APC)或辅助细胞”指呈现与其表面上的主要组织相容性复合体(MHC)复合的外源抗原的细胞。T细胞可使用它们的T细胞受体(TCR)来识别这些复合体。这些细胞加工抗原并且向T细胞呈递它们。专业抗原呈递细胞主要类型:树突细胞(DC)、巨噬细胞、单核细胞及若干B细胞。
术语“抗原呈递细胞(APC)结合域”指能够结合抗原呈递细胞(APC)的区域。所述APC结合域可为多肽,其包含与选自SEQ ID NO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8及SEQ ID NO:9的序列至少90%相同的氨基酸序列。APC结合域为识别并结合在APC上的受体的配体。
分化簇91(CD91)为在细胞膜中形成受体且参与受体介导的内吞作用的蛋白质。
术语“蛋白转导域”指具有致敏T细胞并且因此增强抗原特异性T细胞反应,和/或将抗原导向或指向(即,靶向)抗原呈递的第I类主要组织相容性复合体(MHC-I)通路(即,细胞毒性T细胞通路)的功能的多肽或融合多肽。
术语“致敏T细胞”通常指致敏CD8+及CD4+T细胞,并且因此增强CD8+(CTL)及CD4+T细胞对抗原激发的反应。抗原特异性细胞介导的免疫反应通过在对抗原的反应中定量抗原特异性诱导的γ-干扰素的产生来测量。例如,在无致敏信号(即,无蛋白转导域)的情况下,抗原仅能诱导微弱的细胞介导免疫反应,或根本无法诱导细胞介导的免疫反应,即,CD8+及CD4+T细胞仅制造出少量抗原特异性γ-干扰素,或未制造出抗原特异性γ-干扰素,但在致敏信号(蛋白转导域)存在的情况下,所述抗原可诱导增强的细胞介导的免疫反应。因此,致敏信号(蛋白转导域)的作用在于敏化宿主体内的CD4+及CD8+T细胞,以使得当宿主日后被抗原激发时,由于先前CD4+及CD8+T细胞致敏,所述抗原可可诱导增强的抗原特异性细胞介导的免疫反应。
蛋白转导域可为肽和/或多肽,其选自:
(i)融合多肽,其包含T细胞致敏信号转导肽、连接子及转位肽,其中:
(1)所述T细胞致敏信号转导肽位于所述融合多肽的N端;
(2)所述连接子包含SEQ ID NO:15,其连接所述T细胞致敏信号转导肽与所述转位肽;且
(3)所述转位肽的长度为34-112个氨基酸残基,且包含与SEQ IDNO:3、SEQ ID NO:20或SEQ ID NO:4至少90%相同的氨基酸序列;
(ii)T细胞致敏信号转导肽;及
(iii)长度为34-112个氨基酸残基的转位肽,其包含与SEQ ID NO:3、SEQID NO:20或SEQ ID NO:4至少90%相同的氨基酸序列。
蛋白转导域可为“融合多肽”,其中,所述融合多肽包含T细胞致敏信号转导肽、连接子及转位肽。例如,所述融合多肽可为多肽“CD28convPEt”。
术语“CD28conv”指CD28保守区,其为“T细胞致敏信号转导肽”。其为诱发CD28激动剂抗体的抗原决定簇。
术语“PEt”指长度为34-112个氨基酸残基的转位肽。
“CD28conv”与“PEt”之间存在连接子。融合多肽“CD28convPEt”的方向或排列方式至关重要,因为“CD28conv”(或T细胞致敏信号转导肽)必须位于PEt(或转位肽)的上游,即,PEt必须位于“CD28conv”的C端以获得增强的T细胞反应。“CD28convPEt”与反向融合肽PEtCD28conv相比可很大程度上提高对CD28conv具有特异性的IgG效价(称为CD28特异性激动剂抗体)。所述CD28特异性激动剂抗体可致敏CD4+及CD8+T细胞。方向正确的融合多肽CD28convPEt在CD28conv与PEt区域之间含有连接子(RXRXKR)。所述连接子含有抗原呈递细胞(APC)特异性蛋白酶(组织蛋白酶L)切割位点Lys-Arg(KR)。因此,所述融合蛋白RAP1-CD28conv PEt-抗原-K3可经消化后将形成两个片段:RAP1-CD28conv及PEt-抗原-K3。RAP1-CD28conv片段可在溶酶体中进一步被消化,然后通过MHC II通路向APC细胞表面呈递CD28conv的抗原决定簇,其又诱发产生CD28激动剂抗体的体液免疫反应。因此,CD28激动剂抗体由B细胞产生。所述CD28激动剂抗体可结合T细胞表面上的CD28,并预先活化T细胞(CD4+及CD8+T细胞)。
“T细胞致敏信号转导肽”的长度为28-53个氨基酸残基,且包含与SEQID NO:31至少90%相同的氨基酸序列,其中,Xaa8为I或L;Xaa10为V、F或A;Xaa11为M或L;Xaa17为L或I。
所述T细胞致敏信号转导肽包含关键区K1(I/L)2E3(V/F/A)4(M/L)5Y6P7P8P9Y10(SEQ ID NO:32),其中,(X)2为I或L;(X)4为V、F或A;(X)5为M或L。
以下实施例使用对小鼠具有特异性的T细胞致敏信号转导肽(TDIYFCKIEFMYPPPYLDNEKSNGTIIH,SEQ ID NO:31,其中,X8为I,X10为F,X11为M)。
图7为以融合蛋白RAP1-CD28convPEt-E7-K3为例显示T细胞致敏融合蛋白的作用机理的示意图。RAP1-CD28convPEt-E7-K3自N端至C端依序包含:(1)位于N端的全长RAP1的区域III;(2)CD28conv;(3)连接子;(4)来自假单胞菌外毒素A的修饰的转位肽;(5)全长第16型HPV E7蛋白;及(6)位于C端作为ER滞留信号的三段KDEL。HPV16E7蛋白被加工成用融合蛋白免疫的受试者细胞内的抗原决定簇。相较于仅含有抗原的传统疫苗,RAP1-CD28convPEt-E7-K3可诱发较佳的细胞毒性T细胞(CTL)反应。设计RAP1-CD28convPEt-E7-K3蛋白来改善APC(如树突细胞)的摄取率并且增强向蛋白酶体途径的HPV16E7抗原加工,然后通过MHC I复合体呈递。由疫苗RAP1-CD28convPEt-E7-K3诱发的HPV16E7蛋白特异性CTL免疫反应的作用机理如图7所示:(a)疫苗结合APC(如树突细胞)表面受体(CD91),并经由内吞作用而内在化;(b1)RAP1-CD28convPEt-E7-K3因组织蛋白酶L蛋白酶在转位肽PEt前的位点进行消化而经过蛋白水解;(b2)或RAP1-CD28convPEt-E7-K3循环至E.R.,并且因弗林蛋白酶在转位肽PEt前的位点而经过蛋白水解;(b3)与此同时,RAP1-CD28conv由溶酶体蛋白酶消化,然后通过MHC II向细胞表面呈递CD28conv的抗原决定簇,并且诱发CD28激动剂抗体的产生,这能预先活化T细胞;(c)最重要的步骤是通过转位肽(PEt)使PEt-E7-K3从溶酶体跨膜转位进入细胞质区室;(d)PEt-E7-K3经蛋白酶体途径进行消化,然后通过MHC I复合体呈递E7的抗原决定蔟,并诱发对E7特异性细胞介导的免疫反应。
图8显示不同物种的CD28保守区序列比对及其共有序列。共有序列中画线的序列(KIEVMYPPPY;SEQ ID NO:32,其中,X2为I,X4为V,X5为M)为CD28激动剂抗体识别与结合的关键区。该关键区可以K1(I/L)2E3(V/F/A)4(M/L)5Y6P7P8P9Y10表示,其中,仅在此的第四个氨基酸残基为物种特异性的:并且在人类、大鼠、猪、牛、绵羊、狗及马中应为V;在小鼠中应为F;在火鸡中应为V。所述关键区序列可用K1(X)2E3(X)4(X)5Y6P7P8P9Y10(SEQ ID NO:32)表示,其中,(X)2为I或L;(X)4为V、F或A;(X)5为M或L。
PEt可包含与SEQ ID NO:3或SEQ ID NO:20至少90%相同的氨基酸序列。例如,PEt的氨基酸序列可为PE的a.a.280-a.a.313(SEQ ID NO:3)、a.a.268-a.a.313(SEQ ID NO:20)、a.a.253-a.a.313或a.a.253-a.a.364(SEQID NO:4)。换言之,PEt的氨基酸序列可含有PE区域II(a.a.253至a.a.364;SEQ ID NO:4)的任一区,只要其包含a.a.280-a.a.313(SEQ ID NO:3)必要的片段即可。
抗原可为病原体蛋白、多肽或肽,其可导致由病原体所引发的疾病的原因,或能够在病原体所感染的宿主体内诱发免疫反应;或为在肿瘤细胞内特异性表达的多肽的肿瘤相关抗原(TAA)。所述抗原可选自病原体或癌症细胞,其包括但不局限于人乳头瘤病毒(HPV)、PRRSV、HIV-1、流感病毒、登革热病毒、丙型肝炎病毒(HCV)、乙型肝炎病毒(HBV)、猪环状病毒2(PCV2)、非小细胞肺癌、乳癌、黑色素瘤、淋巴瘤、结肠癌、肝细胞癌,及它们的结合。例如,HPV E7蛋白(E7)、HCV核心蛋白(HCV核心)、HBVX蛋白(HBx)选作开发疫苗的抗原。所述抗原可为两种以上的抗原(选自一或多种病原体蛋白)融合而成的融合抗原。例如,PRRSV ORF6与ORF5的融合抗原,或PRRSV与PCV2病原体的抗原蛋白的融合。
内质网滞留序列的作用在于协助从内吞区室至ER的抗原转位,并将其留在内腔中。内质网滞留序列包含序列Lys Asp Glu Leu(KDEL)或RDEL。ER序列可包含序列KKDLRDELKDEL(SEQ ID NO:16)、KKDELRDELKDEL(SEQ ID NO:17),KKDELRVELKDEL(SEQ ID NO:18),或基本上上由序列KKDLRDELKDEL(SEQ ID NO:16)、KKDELRDELKDEL(SEQ ID NO:17),KKDELRVELKDEL(SEQ ID NO:18)组成,或由序列KKDLRDELKDEL(SEQ ID NO:16)、KKDELRDELKDEL(SEQ ID NO:17),KKDELRVELKDEL(SEQ ID NO:18)组成。
分子量为39kDa(千道尔顿)的受体相关蛋白(RAP1)为ER驻留蛋白,且为低密度脂蛋白受体相关蛋白的分子伴侣(molecular chaperone)。RAP1对CD91具有高结合亲和度(Kd~3nM),且由三个功能类似的区域构成。
本发明涉及通过根据本发明的融合蛋白的T细胞介导的免疫反应的诱导和增强的发现。以RAP1-CD28convPEt-E7-K3为例,RAP1-CD28convPEt-E7-K3疫苗的策略主要致力于刺激T细胞的产生与活化,其能识别的表达靶抗原E7的HPV16受感染细胞。通过传递抗原至树突细胞,能产生抗原特异性CD8+T细胞及CD4+T细胞。第1型辅助CD4+T细胞尤能有效刺激并增强细胞毒性CD8+T细胞的免疫反应。同时,适应性免疫系统的这两个部分具有于身体多个位点杀死HPV16受感染细胞或与HPV16相关的肿瘤细胞而不对正常组织造成显著损伤。
术语“受试者”指人类或非人类动物。
术语“治疗”指对已患有癌症或感染、或有该疾病症状或诱因的有此需要的受试者施用有效量的融合蛋白,藉以治愈,减轻,解除,改善,缓解或预防疾病、其症状或其诱因。基于任一适当诊断方法的结果能够由健康护理专业人士识别该受试者。
术语“有效量”指给予治疗的受试者治疗效果所需的活性化合物的量。如本领域的技术人员所公认的,有效剂量将根据施用的途径、赋形剂的使用以及与其他治疗手段共用的可能性而变化。
缩写:CD 28,分化簇28。
实施例
以下给出根据本发明实施方式的例示性的仪器、设备、方法与它们相关的结果,而不意欲限制本发明的范围。请注意,为读者的便利,标题或子标题可以用于实施例,其不应限制本发明的范围。此外,在此提出或说明某些理论;但是,不论它们正确或错误,它们不应当限制本发明的范围,只要本发明为根据其本身实践,不管任一特定理论或实行方案。
实施例1
表达载体的构建
图1显示融合蛋白RAP1-PE268-313-E7-K3的表达载体,其包含RAP1区域3(SEQ ID NO:5)、转位最小基本肽(PE268-313;SEQ ID NO:20)、抗原E7及内质网滞留序列(K3,或KDEL信号;SEQ ID NO:16、SEQ ID NO:17、SEQ ID NO:18或SEQ ID NO:19)。转位最小基本肽(PE268-313;SEQ ID NO:20)为从a.a.268至a.a.313的PE(SEQ ID NO:10号)多肽序列区获得的。
所述表达载体为利用图10所示的质粒RAP1-K3构建。含有RAP-1区域3与K3的融合基因的质粒RAP1-K3(图10)如下产生:通过PCR法合成编码NdeIRAP1-(EcoRI,XhoI)-K3XhoI的DNA片段,然后利用卡那霉素抗药基因连接到质粒pUC18骨架中获得质粒RAP1-K3(图10)。为产生表达载体RAP1-PE268-313-E7-K3(图1),将编码PE268-313-E7的DNA片段插入质粒RAP1-K3中(图10)。运用类似方法,通过PCR产生编码各种融合肽的DNA片段(图11),并将其分别插入质粒RAP1-K3(图10)中以产生各种融合蛋白的表达载体(图12A-F)。图3显示融合蛋白RAP1-CD28convPEt-E7-K3的表达载体(图3)。
CD28convPEt片段含有组织蛋白酶L及弗林蛋白酶的切割位点。图4显示融合蛋白RAP1-CD28convPEt-E7-K3的氨基酸序列图谱来说明CD28convPEt片段的重要性。两箭头分别指示组织蛋白酶L及弗林蛋白酶的切割位点。这两个切割位点不仅可用做连接CD28conv与PEt-E7的连接子,还允许融合蛋白的切割,以便将PEt-E7-K3片段从溶体释放至细胞质中。由各种病原体引起的任何其他目的抗原可取代E7来与CD28convPEt融合,之后再将所述融合产物插入到图10的质粒中,以形成图11所示的各种融合蛋白的表达载体。
PE268-313的序列为:pletftrhrqprgweqleqcgypvqrlvalylaarlswnqvdqvir(SEQID NO:20)。CD28convPEt的完整序列如下:tdiyfckiefmypppyldneksngtiihrarykrgweqleqcgypvqrlvalylaarlswnqvdqvirgs(SEQID NO:30),画线的序列代表含有组织蛋白酶L与弗林蛋白酶切割位点的连接子序列。
实施例2
蛋白表达
带有蛋白表达载体的大肠杆菌(E.coli)BL21细胞在37℃下,在含有25μg/ml卡那霉素的Luria Bertani培养基中培养。当培养物到达早期对数生长期(A600=0.1至0.4)时,加入异丙基-β-D-硫代半乳糖苷(IPTG),使其最终浓度为0.5至2mM以达诱导的目的。在IPTG诱导4小时后,收集细胞并通过声波震荡来干扰。将含有过表达蛋白的包涵体分离出来,并在8M尿素/TN缓冲液(8M尿素、50mM(Tris)、50mM NaCl,pH 8.0)中溶解。
融合蛋白RAP1-PE268-313-E7-K3的再折叠在4℃下,通过50X体积的TNZ缓冲液(50mM Tris、50mM NaCl及0.01mM ZnCl2,pH 8.0)透析过夜来进行。再折叠的蛋白在还原(包含二硫苏糖醇;+DTT)及非还原(不含二硫苏糖醇;-DTT)条件下经过SDS-PAGE分析(图2)。分析结果显示,大部分再折叠的蛋白在非还原条件下为单体,显示所述RAP1融合蛋白容易再折叠且未聚集(图2)。
图6A为显示表达融合蛋白RAP1-CD28PEt-E7-K3(其含有小鼠CD28conv或人类CD28conv)并从E.coli细胞的包涵体中萃取而出的流程图。SDS-PAGE分析显示,融合蛋白已充分再折叠(图6B)。
图11显示以如下描述的类似方法表达的融合蛋白的列表:(1)RAP1-PE268-313-E7-K3;(2)RAP1-CD28convPEt-E7-K3;(3)RAP1-CD28PEt-E718-K3;(4)RAP1-HCV核心-K3;(5)RAP1-CD28conv-HCV核心-K3;(6)RAP1-CD28convPEt-HCV核心-K3;(7)RAP1-HBx;(8)RAP1-HBx-K3;(9)RAP1-CD28conv-HBx;(10)RAP1-CD28conv-HBx-K3;(11)RAP1-CD28convPEt-HBx;(12)RAP1-CD28convPEt-HBx-K3;(13)RAP1-PCV2ORF2-K3;(14)RAP1-PE268-313-PCV2ORF2-K3;(15)RAP1-CD28convPEt-PCV2ORF2-K3;(16)RAP1-PE268-313-DGD-K3;(17)RAP1-PE268-313-M12-K3;(18)RAP1-PE268-313-PQAB-K3;(19)RAP1-PE268-313-RSAB-K3;(20)RAP1-CD28convPEt-DGD-K3;(21)RAP1-CD28convPEt-M12-K3;(22)RAP1-CD28convPEt-PQAB-K3;(23)RAP1-CD28convPEt-RSAB-K3。使用上述相同的方法再折叠融合蛋白。SDS-PAGE分析的结果显示,这些融合蛋白均已充分再折叠,并且因此用于制备疫苗(图12A-F)。
实施例3
RAP1-PE268-313-E7-K3抑制第16型人乳头瘤病毒(HPV)E7蛋白所诱导的肿瘤生长
如上所述表达所述融合蛋白PE407-E7-K3及RAP1-PE268-313-E7-K3,并通过SDS-PAGE检查蛋白再折叠(图5B)。小鼠通过皮下注射用2x 103个TC-01细胞(带有第16型HPV E7基因的小鼠肺部上皮细胞系)激发,以诱导第16型HPV癌。所述TC-01细胞激发后十二天,小鼠每周一次连续3周经皮下注射接种安慰剂(磷酸盐缓冲溶液+磷酸铝)、PE407-E7-K3(200μg/剂)或RAP1-PE268-313-E7-K3(200μg/剂)(均以AS04C(GlaxoSmithKline)为佐剂)(图5A)。为细胞毒性T淋巴细胞增强佐剂的AS04C包含MPL(单磷酯脂A,免疫增效剂)及磷酸铝(用于抗原传递的蛋白质吸收剂)。术语“K3”代表氨基酸序列KDELKDELKDEL(SEQ ID NO:19)。各组的肿瘤大小及无肿瘤动物数均加以记录(图5C-D)。以AS04C为佐剂的两种疫苗PE407-E7-K3及RAP1-PE268-313-E7-K3显著抑制肿瘤生长。但是,接种RAP1-PE268-313-E7-K3的小鼠组具有较高比例的无肿瘤小鼠。其显示疫苗RAP1-PE268-313-E7-K3在抑制肿瘤生长方面与PE407-E7-K3一样有效,或更为有效,而在提高无肿瘤动物的比例方面更好。
实施例4
RAP1-CD28convPEt-E7-K3可抑制第16型人乳头瘤病毒(HPV)E7蛋白所诱导的肿瘤生长并提高存活率
检验有或没有免疫增效剂的融合蛋白PE407-E7-K3及RAP1-CD28convPEt-E7-K3对肿瘤大小及存活率的影响。小鼠经皮下注射以较高剂量(3x104)的TC-01细胞激发。激发后七天,小鼠以每周一次连续3周经皮下注射接种有免疫增效剂GPI-0100或蛋白质吸收剂磷酸铝的安慰剂、PE407-E7-K3(100μg/剂)或RAP1-CD28convPEt-E7-K3(100μg/剂)(图9A)。GPI-0100为Th1/CTL刺激佐剂(免疫增效剂)。各组的肿瘤大小及存活率均加以记录(图9B-C)。当与佐剂GPI-0100结合时,PE407-E7-K3及RAP1-CD28convPEt-E7-K3两者抑制肿瘤生长。意外地,发现RAP1-CD28convPEt-E7-K3抑制肿瘤生长的效果不取决于佐剂。当与所述蛋白质吸收剂磷酸铝结合而未与佐剂GPI-0100结合时,RAP1-CD28convPEt-E7-K3仍可以与免疫增效剂GPI-0100结合时的相同效力有效抑制肿瘤生长(图9B,空心三角形对实心反向三角形)。
相比之下,PE407-E7-K3抑制肿瘤生长的效力取决于佐剂。当与蛋白质吸收剂磷酸铝结合时,PE407-E7-K3的效力低于与所述免疫增效剂GPI-0100结合时的效力(图9B,实心正方形对空心圆形)。
另一方面,就存活率而言,施用与所述免疫增效剂GPI-0100或所述蛋白质吸收剂磷酸铝结合的RAP1-CD28convPEt-E7-K3的小鼠具有比接种与GPI-0100或磷酸铝结合的PE407-E7-K3更高的存活率(图9C)。其显示,RAP1-CD28convPEt-E7-K3即使无免疫增效剂GPI-0100,仍可诱发Th1/CTL免疫反应。所述结果亦显示,作为用于提高动物存活率的疫苗,所述融合蛋白RAP1-CD28convPEt-E7-K3优于PE407-E7-K3。
实施例5
免疫原性分析
测试多种疫苗的免疫原性。小鼠分为下列各组:HPV16E7、HPV 18E7、HCV核心、HBV HBx、PCV2ORF2及PRRSV(图13A),各组再划分为数个亚组,各亚组依每周一次连续3周经皮下注射安慰剂或针对病原体的一种或多种抗原而设计的疫苗(图13B)。针对PRRSV的疫苗除外,各个疫苗由单一融合蛋白及吸收剂磷酸铝组成,其中,所述单一融合蛋白含有病原体的至少一种抗原。所述抗原为病原体的全长蛋白,或为含有病原体的抗原的至少一种抗原决定簇的非全长蛋白,或为两种以上抗原的融合肽,其中,各抗原为选自病原体的不同蛋白。
免疫接种程序、疫苗及剂量在图13A-B中显示。简言之,小鼠每周一次连续3周接种图13A列出的疫苗。所有小鼠均在最后一次免疫7后天处死,并收集其脾脏。脾细胞经分离后,在有10μg/ml的各种病原体重组抗原的6孔培养板(2x107个细胞/2ml/孔)中培养,以在37℃下在1μg/ml GolgiPlug(BD Pharmingen,San Diego,CA)存在下刺激脾细胞16小时。
刺激后的脾细胞以FACScan缓冲液清洗,并且所述细胞表面标记CD8a、CD4及CD3则以藻红蛋白共轭的单克隆大鼠抗小鼠CD8a抗体、AF700共轭的单克隆大鼠抗小鼠CD4抗体及AF647共轭的单克隆大鼠抗小鼠CD3抗体染色。接着再以根据制造商的说明(BD Pharmingen)的Cytofix/Cytoperm套组浸透并固定细胞。细胞内的IFN-γ为以AF488共轭的单克隆大鼠抗小鼠IFN-γ染色,以便量测免疫反应及细胞因子水平。使用Gallios流式细胞术及Kaluza分析软件(Beckman Coulter)进行流式细胞术分析。
测试下列PRRSV疫苗的免疫原性:PE407-PRRSV-K3、RAP1-PE268-313-PRRSV-K3或RAP1-CD28convPEt-PRRSV-K3疫苗。各疫苗含有四种不同融合蛋白的混合物,且各融合蛋白含有选自DGD、M12、PQAB及RSAB的不同抗原(图13A)。小鼠的接种及脾细胞的刺激使用与上述类似的方式进行。简言之,所有小鼠在最后一次免疫后7天处死,并收集脾脏。分离脾细胞,并且分别在有10μg/ml的重组DGD、M12、PQAB或RSAB抗原的6孔培养板(2x107个细胞/2ml/孔)中培养,以在37℃下及1μg/mlGolgiPlug(BD Pharmingen,San Diego,CA)存在下刺激脾细胞16小时。
“DGD”(SEQ ID NO:26)的氨基酸序列如下:
ALDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAES ERFVRQGTGNDEAGAANADVVSLTCPVAAGECAGPADSGDALLERNYPT GAEFLGDGGDV DGD代表PRRSV ORF7a.a.64-a.a.123(粗体)、连接子(画线)及ORF7a.a.64-a.a.123(粗体)的融合抗原。
术语“M12”代表PRRSV ORF1b a.a.1046-a.a.1210的抗原。其氨基酸序列(SEQ ID NO:27)如下:NNKECTVAQALGNGDKFRATDKRVVDSLRAICADLEGSSSPLPKVAHNLGFYFSPDLTQFAKLPIELAPHWPVVSTQNNEKWPDRLVASLRPLDKYSRACIGAGYMVGPSVFLGTPGVVSYYLTKFVKGEAQVLPETVFSTGRIEVDCREYLDDREREVAASLPH。
“PQAB”(SEQ ID NO:28)的氨基酸序列如下:GSSLDDFCYDSTAPQKVLLAFSITYASNDSSSHLQLIYNLTLCELNGTDWL ANKFDWA。PQAB代表PRRSV美洲株ORF6a.a.2-a.a.26及ORF5a.a.31-a.a.63(画线)的融合抗原。
RSAB的氨基酸序列为MGSLDDFCNDSTAAQKLVLAFSITYTPIFVAGGSSSTYQYIYNLTICELNGT DWLSNHFDWA(SEQ ID NO:29)。术语“RSAB”代表PRRSV欧洲株ORF6a.a.2-28及ORF5a.a.31-64(画线)的融合抗原。
实施例6
融合蛋白RAP1-CD28convPEt-E7-K3的RAP1区域3的片段分别由A2M最小(SEQ ID NO:6)、HIV-Tat最小(SEQ ID NO:7)或HSPs最小(SEQ IDNO:8)取代,以产生融合蛋白A2M-CD28convPEt-E7-K3、Tat-CD28convPEt-E7-K3及HSP-CD28convPEt-E7-K3疫苗。使用与上述类似的方法检测由这些疫苗增强的TC-1肿瘤抑制活性及细胞介导的免疫反应。表1显示各融合蛋白的组分的序列编号(SEQ ID NO)。表2显示测试融合蛋白对动物T细胞介导的免疫反应的影响的及其抗原序列。
表1
表2
在免疫原性测定中,通过测量脾细胞中CD3+/CD4+/IFNγ+及CD3+/CD8+/IFNγ+T细胞的数量来评估由各疫苗所诱导的抗原特异性细胞介导的免疫反应。结果显示,所述疫苗RAP1-CD28convPEt-抗原-K3可诱导强烈的T细胞反应。图14B显示,由CD28convPEt-E718-K3所诱发的CD3+/CD4+/IFNγ+T细胞数为RAP1-K3所诱发的细胞数的大约50倍,由CD28convPEt-E718-K3所诱发的CD3+/CD8+/IFNγ+T细胞数则为RAP1-K3所诱发的细胞数的9倍以上。
疫苗RAP1-CD28convPEt-抗原-K3在诱发T细胞介导的免疫原性方面优于PE407-抗原-K3。例如,图14A显示,由CD28convPEt-E716-K3所诱发的CD3+/CD4+/IFNγ+T细胞数及CD3+/CD8+/IFNγ+T细胞数分别为PE407-E716-K3所诱发的T细胞数的大约5倍与7倍。其显示,所述疫苗RAP1-CD28convPEt-E7-K3具有比PE407-E7-K3更好的细胞介导的免疫原性。
当选用的抗原包含10个或多于10个抗原决定簇时,包含RAP1区域III、致敏信号CD28conv(但不含转位肽PEt)、抗原及ER滞留信号的融合蛋白便足以诱发强烈的抗原特异性T细胞介导的免疫反应。图14C显示疫苗RAP1-CD28conv-HCV核心-K3诱发CD3+/CD4+/IFNγ+及CD3+/CD8+/IFNγ+T细胞的数量分别为安慰剂组的20倍及7.6倍的T细胞反应。抗原HCV核心含有11个众所周知的MHC I抗原决定簇。
意外地,所述ER滞留信号对于本发明的融合蛋白诱发强烈的细胞介导的免疫原性来说不是必要的。换言之,在ER滞留序列不存在的情况下,本发明的融合蛋白仍可诱发强烈的T细胞反应。图14D显示,由RAP1-CD28convPEt-HBx(其不含ER滞留信号K3)所诱发的CD3+/CD4+/IFNγ+及CD3+/CD8+/IFNγ+T细胞的数量为安慰剂组的7倍及74倍。
相较之下,美国专利第7378100B2号及第7335361号显示所述ER滞留信号K3对PE相关的融合蛋白(PE407-抗原-K3)诱发T细胞反应来说是必须的。
还发现,包含RAP1区域III、转位肽PE218-313(但不含致敏信号CD28conv)、抗原及ER滞留信号的融合蛋白优于不含RAP1区域III的PE相关的融合蛋白。图15C-J显示,所述疫苗RAP1-PE268-313-PRRSV-K3所诱发的CD3+/CD4+/IFNγ+及CD3+/CD8+/IFNγ+T细胞数大于所述疫苗PE407-PRRSV-K3所诱发的T细胞数。
已介绍上述的本发明的示例性实施方式仅用于说明和描述的目的,并未意欲成为全面的,或意欲限制本发明至说明的精确形式。依据以上教示可实现本发明的多种修改及变化。选择并描述实施方式与实施例以便解释本发明的原理与它们的实际应用,为了使本领域的其他技术人员能够利用本发明及各种实施方式及适合预期的特定使用的各种修改。在不偏离本发明的实质和范围的情况下,本发明有关的选择性的实施方式将对本领域的这些技术人员明显可见。因此,本发明的范围由所附权利要求来界定,而非由以上说明和在此描述的示例性实施方式来界定。
Claims (20)
1.一种融合蛋白,其包含:
(a)抗原呈递细胞(APC)结合域或CD91受体结合域,其位于所述融合蛋白的N端;
(b)蛋白转导域,其位于所述APC结合域或CD91受体结合域的C端,所述蛋白转导域选自:
(i)融合多肽,其包含T细胞致敏信号转导肽、连接子及转位肽,其中:
(1)所述T细胞致敏信号转导肽位于所述融合多肽的N端;
(2)所述连接子包含SEQ ID NO:15,其连接所述T细胞致敏信号转导肽与所述转位肽;且
(3)所述转位肽的长度为34-112个氨基酸残基,且包含与SEQID NO:3、SEQ ID NO:20或SEQ ID NO:4至少90%相同的氨基酸序列;
(ii)T细胞致敏信号转导肽;及
(iii)长度为34-112个氨基酸残基的转位肽,其包含与SEQ ID NO:3、SEQ ID NO:20或SEQ ID NO:4至少90%相同的氨基酸序列;及
(c)病原体的抗原,其位于所述蛋白转导域的C端;
其中:
所述T细胞致敏信号转导肽的长度为28-53个氨基酸残基,且包含与SEQID NO:31至少90%相同的氨基酸序列,其中,Xaa8为I或L;Xaa10为V、F或A;Xaa11为M或L;Xaa17为L或I;且
若所述蛋白转导域为转位肽(biii),则所述APC结合域或CD91受体结合域无假单胞菌外毒素A(PE)结合域I的氨基酸序列。
2.如权利要求1所述的融合蛋白,其中,所述APC结合域或CD91受体结合域为包含与选自SEQ ID NO:5、SEQ ID NO:9、SEQ ID NO:6、SEQ IDNO:7及SEQ ID NO:8的序列至少90%相同的氨基酸序列的多肽。
3.如权利要求1所述的融合蛋白,进一步包含位于所述融合蛋白的C端的内质网滞留序列。
4.如权利要求3所述的融合蛋白,其中,所述内质网滞留序列包含氨基酸序列Lys-Asp-Glu-Leu(SEQ ID NO:14)。
5.如权利要求1所述的融合蛋白,其中,若所述抗原含有10个或更多抗原决定簇,则所述融合蛋白的C端无内质网滞留序列。
6.如权利要求1所述的融合蛋白,其中,所述蛋白转导域为所述融合多肽(bi)。
7.如权利要求1所述的融合蛋白,其中,所述蛋白转导域为所述T细胞致敏信号转导肽(bii)。
8.如权利要求7所述的融合蛋白,其在所述蛋白转导域与所述抗原之间进一步包含另一连接子,所述另一连接子包含SEQ ID NO:15。
9.如权利要求1所述的融合蛋白,其中,所述蛋白转导域为转位肽(biii)。
10.如权利要求9所述的融合蛋白,其在所述APC结合域或CD91受体结合域与所述转位肽之间进一步包含另一连接子,所述另一连接子包含SEQID NO:15。
11.如权利要求1所述的融合蛋白,其中,所述蛋白转导域包含序列SEQID NO:30。
12.一种疫苗组合物,其包含:
(a)治疗有效量的权利要求1所述的融合蛋白;及
(b)佐剂。
13.如权利要求1所述的融合蛋白,其中,所述APC结合域或CD91受体结合域为包含选自SEQ ID NO:5、SEQ ID NO:9、SEQ ID NO:6、SEQ IDNO:7及SEQ ID NO:8的氨基酸序列的多肽。
14.如权利要求1所述的融合蛋白,其中,所述T细胞致敏信号转导肽包含与SEQ ID NO:1或SEQ ID NO:2至少90%相同的氨基酸序列。
15.如权利要求14所述的融合蛋白,其中,所述T细胞致敏信号转导肽包含选自SEQ ID NO:1及SEQ ID NO:2的氨基酸序列。
16.如权利要求1所述的融合蛋白,其中,所述转位肽包含选自SEQ IDNO:3、SEQ ID NO:20及SEQ ID NO:4的氨基酸序列。
17.如上述权利要求中的任意一项所述的融合蛋白或疫苗组合物,其用于诱导增强的病原体抗原特异性T细胞反应。
18.如上述权利要求中的任意一项所述的融合蛋白或疫苗组合物,其用于杀死通过在疾病细胞的细胞膜上的第I类主要组织相容性复合体(MHC)分子呈递抗原的疾病细胞。
19.如权利要求18所述使用的融合蛋白或疫苗组合物,其中,所述疾病细胞为癌细胞。
20.如上述权利要求中的任意一项所述的融合蛋白或疫苗组合物,其用于预防、治疗由病原体所引起的感染和/或将由感染所引起的症状减至最少。
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201811288421.3A CN109535228B (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811287074.2A CN109535259A (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811287071.9A CN109553688A (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811287073.8A CN109608550B (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201261733879P | 2012-12-05 | 2012-12-05 | |
US61/733,879 | 2012-12-05 | ||
PCT/US2013/072753 WO2014089009A1 (en) | 2012-12-05 | 2013-12-03 | Fusion proteins for use as immunogenic enhancers for inducing antigen-specific t cell responses |
Related Child Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201811288421.3A Division CN109535228B (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811287074.2A Division CN109535259A (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811287073.8A Division CN109608550B (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811287071.9A Division CN109553688A (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN104918637A true CN104918637A (zh) | 2015-09-16 |
CN104918637B CN104918637B (zh) | 2018-12-11 |
Family
ID=50825669
Family Applications (7)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201811287071.9A Pending CN109553688A (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201380063896.1A Expired - Fee Related CN104918637B (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201710718246.6A Expired - Fee Related CN107434827B (zh) | 2012-12-05 | 2013-12-03 | 用作诱导抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811287073.8A Expired - Fee Related CN109608550B (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811287074.2A Pending CN109535259A (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811288421.3A Expired - Fee Related CN109535228B (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201380063867.5A Expired - Fee Related CN104884082B (zh) | 2012-12-05 | 2013-12-03 | 用作诱导抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201811287071.9A Pending CN109553688A (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
Family Applications After (5)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201710718246.6A Expired - Fee Related CN107434827B (zh) | 2012-12-05 | 2013-12-03 | 用作诱导抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811287073.8A Expired - Fee Related CN109608550B (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811287074.2A Pending CN109535259A (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201811288421.3A Expired - Fee Related CN109535228B (zh) | 2012-12-05 | 2013-12-03 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN201380063867.5A Expired - Fee Related CN104884082B (zh) | 2012-12-05 | 2013-12-03 | 用作诱导抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
Country Status (12)
Country | Link |
---|---|
US (4) | US9481714B2 (zh) |
EP (2) | EP2928491B1 (zh) |
JP (4) | JP6173479B2 (zh) |
KR (3) | KR101746444B1 (zh) |
CN (7) | CN109553688A (zh) |
BR (2) | BR112015013135A2 (zh) |
DK (1) | DK2928491T3 (zh) |
ES (2) | ES2742739T3 (zh) |
IL (2) | IL239158A0 (zh) |
RU (2) | RU2619187C2 (zh) |
TW (2) | TWI490231B (zh) |
WO (2) | WO2014089036A1 (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107847575A (zh) * | 2015-06-01 | 2018-03-27 | 瑞宝基因股份有限公司 | 用于猪生殖与呼吸综合症及猪圆环病毒相关疾病的疫苗组合物 |
CN109715651A (zh) * | 2016-09-19 | 2019-05-03 | 生控基因疫苗股份有限公司 | 乙型肝炎治疗型疫苗 |
Families Citing this family (22)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11246915B2 (en) | 2010-09-15 | 2022-02-15 | Applied Molecular Transport Inc. | Cholix toxin-derived fusion molecules for oral delivery of biologically active cargo |
CN109553688A (zh) * | 2012-12-05 | 2019-04-02 | 生控基因疫苗股份有限公司 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
CN107249629A (zh) * | 2015-02-26 | 2017-10-13 | 生控基因疫苗股份有限公司 | 包含免疫原性蛋白质及组合佐剂并用以诱发抗原特异性t细胞反应的疫苗组合物 |
JP2018521983A (ja) | 2015-07-16 | 2018-08-09 | バイオカイン セラピューティックス リミテッド | がんを治療するための組成物および方法 |
KR102153303B1 (ko) * | 2015-11-23 | 2020-09-09 | 뵈링거 잉겔하임 애니멀 헬스 유에스에이 인코포레이티드 | Fmdv 및 e2 융합 단백질 및 이의 용도 |
US11186618B2 (en) * | 2016-09-21 | 2021-11-30 | The Research Foundation For Microbial Diseases Of Osaka University | Dendritic-cell-targeted peptide, fusion peptide utilizing said peptide, and vaccine utilizing said fusion peptide |
CN107991481A (zh) * | 2016-10-27 | 2018-05-04 | 武汉科前生物股份有限公司 | 一种检测猪伪狂犬病毒和口蹄疫病毒的二联阻断elisa抗体检测试剂盒及其应用 |
CN107937354A (zh) * | 2017-11-10 | 2018-04-20 | 南京天邦生物科技有限公司 | 2型猪圆环病毒及其应用 |
PL3762009T3 (pl) | 2018-03-08 | 2022-09-12 | Applied Molecular Transport Inc. | Pochodzące z toksyny konstrukty dostarczające do dostarczania doustnego |
CN109134667A (zh) * | 2018-09-19 | 2019-01-04 | 天康生物股份有限公司 | 融合蛋白及其制备方法、应用、表达系统和疫苗 |
JP2022512976A (ja) | 2018-11-07 | 2022-02-07 | アプライド モレキュラー トランスポート インコーポレイテッド | 異種ペイロードの経口送達のためのコリックス由来担体 |
CN109593136B (zh) * | 2018-12-26 | 2021-03-19 | 天康生物股份有限公司 | 禽副粘病毒融合蛋白及其制备方法、应用和用于鸽子的apmv疫苗 |
KR20220012256A (ko) * | 2019-05-24 | 2022-02-03 | 프로비바 테라퓨틱스 (홍콩) 리미티드 | Il-2 조성물 및 이의 사용 방법 |
CN110051832B (zh) * | 2019-05-31 | 2020-11-10 | 四川农业大学 | 一种犬恶丝虫病疫苗 |
CN110452928A (zh) * | 2019-08-13 | 2019-11-15 | 成都天邦生物制品有限公司 | 一株pk-15稳定细胞株的构建及应用 |
AU2020329290A1 (en) * | 2019-08-13 | 2022-03-24 | Elpis Biopharmaceuticals | Engineered interleukin-2 receptor beta agonists |
CN110669142B (zh) * | 2019-09-30 | 2021-10-12 | 湖南农业大学 | 融合rgd的猪圆环病毒2型病毒样颗粒、突变型感染性克隆及其制备方法和应用 |
US20230173049A1 (en) * | 2019-12-31 | 2023-06-08 | The Johns Hopkins University | Fusion proteins and methods of use thereof |
CN111548395A (zh) * | 2020-05-25 | 2020-08-18 | 中国农业科学院兰州兽医研究所 | 一种口蹄疫病毒二价多表位重组病毒样颗粒及其应用 |
KR102507298B1 (ko) | 2020-12-21 | 2023-03-08 | 충남대학교산학협력단 | 면역증강을 위한 조성물 |
CN112812171B (zh) * | 2021-01-22 | 2022-04-08 | 浙江辉肽生命健康科技有限公司 | 具有氨基酸结构vvrkplnkegkkp的生物活性肽及其制备方法和应用 |
CN114539429B (zh) * | 2022-03-24 | 2022-07-29 | 上海普铭生物科技有限公司 | 融合蛋白组合物及其应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5686281A (en) * | 1995-02-03 | 1997-11-11 | Cell Genesys, Inc. | Chimeric receptor molecules for delivery of co-stimulatory signals |
WO2004087196A2 (en) * | 2003-04-03 | 2004-10-14 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Methods and compositions for the inhibition of modulation of t cell costimulatory pathway by a pathogenic agent |
US20090214570A1 (en) * | 2005-05-18 | 2009-08-27 | Trinity Biosystems, Inc. | Methods and compositions for immunizing against chlamydia infection |
Family Cites Families (36)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US843505A (en) * | 1905-10-09 | 1907-02-05 | Alexandre Leonard Tombelaine | Safety closing device for miner's lamps. |
PT651805E (pt) * | 1992-07-17 | 2007-02-28 | Dana Farber Cancer Inst Inc | Método de ligação intracelular de moléculas-alvo |
US6451592B1 (en) * | 1996-04-05 | 2002-09-17 | Chiron Corporation | Recombinant alphavirus-based vectors with reduced inhibition of cellular macromolecular synthesis |
US20050119470A1 (en) * | 1996-06-06 | 2005-06-02 | Muthiah Manoharan | Conjugated oligomeric compounds and their use in gene modulation |
DE19651443A1 (de) * | 1996-12-11 | 1998-06-18 | Hoechst Ag | Selbstverstärkende, pharmakologisch kontrollierbare Expressionssysteme |
US7314632B1 (en) | 1997-07-11 | 2008-01-01 | The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services | Pseudomonas exotoxin A-like chimeric immunogens |
DK1000163T3 (da) * | 1997-07-11 | 2006-02-06 | Us Gov Health & Human Serv | Pseudomonas-exotoksin A-lignende kimære immunogener |
ATE359084T1 (de) * | 1998-02-20 | 2007-05-15 | Univ Miami | Modifizierter hitzeschockprotein/peptidantigen komplex |
WO1999045127A2 (en) * | 1998-03-06 | 1999-09-10 | Oxford Biomedica (Uk) Limited | Enhanced prodrug activation |
US20030215421A1 (en) * | 1999-07-21 | 2003-11-20 | Mcdonald John R. | Methods and compositions for treating secondary tissue damage and other inflammatory conditions and disorders |
US20020061848A1 (en) * | 2000-07-20 | 2002-05-23 | Ajay Bhatia | Compounds and methods for treatment and diagnosis of chlamydial infection |
WO2000049041A1 (fr) * | 1999-02-19 | 2000-08-24 | Sumitomo Electric Industries, Ltd. | Preparations proteiques |
JP4637368B2 (ja) * | 1999-04-14 | 2011-02-23 | ノバルティス バクシンズ アンド ダイアグノスティックス,インコーポレーテッド | アルファウイルスに基づくベクター系を利用する免疫応答を生成するための組成物および方法 |
EP1222289B1 (en) * | 1999-10-20 | 2008-04-16 | The Johns Hopkins University School Of Medicine | Chimeric immunogenic compositions and nucleic acids encoding them |
US8128922B2 (en) * | 1999-10-20 | 2012-03-06 | Johns Hopkins University | Superior molecular vaccine linking the translocation domain of a bacterial toxin to an antigen |
US7030219B2 (en) * | 2000-04-28 | 2006-04-18 | Johns Hopkins University | B7-DC, Dendritic cell co-stimulatory molecules |
CA2453528C (en) * | 2001-08-01 | 2011-07-26 | Cellomics, Inc. | Novel fusion proteins and assays for molecular binding |
US7235631B2 (en) * | 2002-02-07 | 2007-06-26 | Mayo Foundation For Medical Education And Research | ICOS mutants |
IL164799A0 (en) * | 2002-04-25 | 2005-12-18 | Univ Connecticut | Using heat shock proteins to improve the therapeutic benefit of a non-vaccine treatment modality |
CA2515123A1 (en) * | 2003-02-04 | 2004-08-19 | University Of Connecticut Health Center | Immunogenic cd91 ligand-antigenic molecule complexes and fusion proteins |
EP1594437A2 (en) * | 2003-02-04 | 2005-11-16 | University of Connecticut Health Center | Immunogenic cd91 ligand-antigenic molecule complexes and fusion proteins |
US7595054B2 (en) * | 2003-06-09 | 2009-09-29 | Healthbanks Biotech Co., Ltd. | Fusion antigen used as vaccine |
US7335361B2 (en) | 2003-06-09 | 2008-02-26 | Animal Technology Institute Taiwan | Fusion antigen used as vaccine |
EP1756147A2 (en) * | 2004-06-01 | 2007-02-28 | Innogenetics N.V. | Peptides for inducing a ctl and/or htl response to hepatitis c virus |
JP4474264B2 (ja) * | 2004-08-20 | 2010-06-02 | 生寶生物科技股▲ふん▼有限公司 | 子宮頸癌抑制の融合蛋白 |
WO2006135428A2 (en) * | 2004-10-04 | 2006-12-21 | Trinity Biosystems, Inc. | Methods and compositions for inducing an immune response against multiple antigens |
US7465455B2 (en) | 2006-07-05 | 2008-12-16 | Healthbanks Biotech Co., Ltd. | Fusion protein of porcine reproductive and respiratory syndrome virus as PRRS vaccine |
WO2008047243A2 (en) * | 2006-08-29 | 2008-04-24 | Forhumantech. Co., Ltd. | Pharmaceutical composition for suppression of apoptosis and method for delivering the same |
MX2009002893A (es) * | 2006-09-18 | 2009-07-10 | Raptor Pharmaceutical Inc | Tratamiento de trastornos hepaticos mediante la administracion de conjugados de la proteina asociada al receptor (rap). |
US7887801B2 (en) * | 2007-07-13 | 2011-02-15 | Topotarget Germany Ag | Optimized DNA and protein sequence of an antibody to improve quality and yield of bacterially expressed antibody fusion proteins |
WO2009036349A1 (en) * | 2007-09-12 | 2009-03-19 | Anaphore, Inc. | Hsp70-based treatment for autoimmune diseases |
EP2078726A1 (en) * | 2008-01-09 | 2009-07-15 | Vision 7 GmbH | Secretable HIV entry inhibitory peptides for therapy of HIV infection |
CN101969975B (zh) * | 2008-01-31 | 2013-06-05 | 生控基因疫苗股份有限公司 | 作为疫苗的hiv嵌合融合蛋白 |
WO2010040023A2 (en) * | 2008-10-03 | 2010-04-08 | Government Of The United States Of America, As Represented By The Secretary, Department Of Health & Human Services | Methods and compositions for protein delivery |
PL391627A1 (pl) * | 2010-06-25 | 2012-01-02 | Adamed Spółka Z Ograniczoną Odpowiedzialnością | Przeciwnowotworowe białko fuzyjne |
CN109553688A (zh) * | 2012-12-05 | 2019-04-02 | 生控基因疫苗股份有限公司 | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 |
-
2013
- 2013-12-03 CN CN201811287071.9A patent/CN109553688A/zh active Pending
- 2013-12-03 CN CN201380063896.1A patent/CN104918637B/zh not_active Expired - Fee Related
- 2013-12-03 ES ES13860021T patent/ES2742739T3/es active Active
- 2013-12-03 KR KR1020167029734A patent/KR101746444B1/ko active IP Right Grant
- 2013-12-03 CN CN201710718246.6A patent/CN107434827B/zh not_active Expired - Fee Related
- 2013-12-03 WO PCT/US2013/072804 patent/WO2014089036A1/en active Application Filing
- 2013-12-03 BR BR112015013135A patent/BR112015013135A2/pt not_active Application Discontinuation
- 2013-12-03 US US14/095,760 patent/US9481714B2/en active Active
- 2013-12-03 US US14/095,947 patent/US9339536B2/en active Active
- 2013-12-03 JP JP2015546553A patent/JP6173479B2/ja not_active Expired - Fee Related
- 2013-12-03 EP EP13860917.7A patent/EP2928491B1/en not_active Not-in-force
- 2013-12-03 ES ES13860917T patent/ES2701448T3/es active Active
- 2013-12-03 CN CN201811287073.8A patent/CN109608550B/zh not_active Expired - Fee Related
- 2013-12-03 WO PCT/US2013/072753 patent/WO2014089009A1/en active Application Filing
- 2013-12-03 RU RU2015122368A patent/RU2619187C2/ru not_active IP Right Cessation
- 2013-12-03 CN CN201811287074.2A patent/CN109535259A/zh active Pending
- 2013-12-03 BR BR112015013183A patent/BR112015013183A2/pt not_active Application Discontinuation
- 2013-12-03 CN CN201811288421.3A patent/CN109535228B/zh not_active Expired - Fee Related
- 2013-12-03 DK DK13860917.7T patent/DK2928491T3/en active
- 2013-12-03 JP JP2015546546A patent/JP6279606B2/ja not_active Expired - Fee Related
- 2013-12-03 KR KR1020157014330A patent/KR101650364B1/ko active IP Right Grant
- 2013-12-03 CN CN201380063867.5A patent/CN104884082B/zh not_active Expired - Fee Related
- 2013-12-03 RU RU2015122371A patent/RU2631002C2/ru not_active IP Right Cessation
- 2013-12-03 KR KR1020157014327A patent/KR101671291B1/ko active IP Right Grant
- 2013-12-03 EP EP13860021.8A patent/EP2928493B1/en not_active Not-in-force
- 2013-12-05 TW TW102144581A patent/TWI490231B/zh active
- 2013-12-05 TW TW102144580A patent/TWI486359B/zh active
-
2015
- 2015-06-03 IL IL239158A patent/IL239158A0/en unknown
- 2015-06-03 IL IL239157A patent/IL239157A0/en unknown
-
2016
- 2016-04-13 US US15/098,210 patent/US9676826B2/en not_active Expired - Fee Related
- 2016-09-22 US US15/273,588 patent/US9676827B2/en active Active
-
2017
- 2017-07-04 JP JP2017130836A patent/JP6449384B2/ja not_active Expired - Fee Related
- 2017-08-30 JP JP2017166010A patent/JP6400810B2/ja not_active Expired - Fee Related
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5686281A (en) * | 1995-02-03 | 1997-11-11 | Cell Genesys, Inc. | Chimeric receptor molecules for delivery of co-stimulatory signals |
WO2004087196A2 (en) * | 2003-04-03 | 2004-10-14 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Methods and compositions for the inhibition of modulation of t cell costimulatory pathway by a pathogenic agent |
US20090214570A1 (en) * | 2005-05-18 | 2009-08-27 | Trinity Biosystems, Inc. | Methods and compositions for immunizing against chlamydia infection |
Non-Patent Citations (2)
Title |
---|
CHIANG等: "Novel Synthetic Bipartite Carrier Protein for Developing Glycotope-Based Vaccines", 《VACCINE 》 * |
TAKEMOTO等: "Enhanced Generation of Cytotoxic T Lymphocytes by Heat Shock Protein 70 Fusion Proteins Harboring Both CDB+ T Cell and CD4+ T Cell Epitopes", 《MOL PHARM》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107847575A (zh) * | 2015-06-01 | 2018-03-27 | 瑞宝基因股份有限公司 | 用于猪生殖与呼吸综合症及猪圆环病毒相关疾病的疫苗组合物 |
CN107847575B (zh) * | 2015-06-01 | 2021-12-17 | 瑞宝基因股份有限公司 | 用于猪生殖与呼吸综合症及猪圆环病毒相关疾病的疫苗组合物 |
CN109715651A (zh) * | 2016-09-19 | 2019-05-03 | 生控基因疫苗股份有限公司 | 乙型肝炎治疗型疫苗 |
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN104918637A (zh) | 用作诱发抗原特异性t细胞反应的免疫原性增强剂的融合蛋白 | |
US20160250324A1 (en) | Vaccine composition comprising an immunogenic protein and combination adjuvants for use in eliciting antigen-specific t-cell responses | |
CN102813918B (zh) | 一种猪圆环病毒2型重组亚单位疫苗及其制备方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20181211 Termination date: 20201203 |