CN117264066A - 一种eno1抗体及其应用 - Google Patents
一种eno1抗体及其应用 Download PDFInfo
- Publication number
- CN117264066A CN117264066A CN202310510154.4A CN202310510154A CN117264066A CN 117264066 A CN117264066 A CN 117264066A CN 202310510154 A CN202310510154 A CN 202310510154A CN 117264066 A CN117264066 A CN 117264066A
- Authority
- CN
- China
- Prior art keywords
- seq
- monoclonal antibody
- amino acid
- nucleic acid
- antibody
- 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
- 102100038910 Alpha-enolase Human genes 0.000 title claims abstract description 47
- 101000882335 Homo sapiens Alpha-enolase Proteins 0.000 title claims abstract description 45
- VLMZMRDOMOGGFA-WDBKCZKBSA-N festuclavine Chemical compound C1=CC([C@H]2C[C@H](CN(C)[C@@H]2C2)C)=C3C2=CNC3=C1 VLMZMRDOMOGGFA-WDBKCZKBSA-N 0.000 title claims abstract description 38
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 45
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 40
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 38
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 38
- 239000013598 vector Substances 0.000 claims abstract description 20
- 201000008482 osteoarthritis Diseases 0.000 claims abstract description 8
- 208000001132 Osteoporosis Diseases 0.000 claims abstract description 7
- 206010039073 rheumatoid arthritis Diseases 0.000 claims abstract description 7
- 210000004027 cell Anatomy 0.000 claims description 60
- 239000003814 drug Substances 0.000 claims description 21
- 239000002773 nucleotide Substances 0.000 claims description 20
- 125000003729 nucleotide group Chemical group 0.000 claims description 20
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 18
- 238000011282 treatment Methods 0.000 claims description 18
- 239000008194 pharmaceutical composition Substances 0.000 claims description 17
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 16
- 239000000126 substance Substances 0.000 claims description 16
- 238000006467 substitution reaction Methods 0.000 claims description 16
- 101150015836 ENO1 gene Proteins 0.000 claims description 15
- 239000003153 chemical reaction reagent Substances 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 14
- 201000010099 disease Diseases 0.000 claims description 13
- 229940079593 drug Drugs 0.000 claims description 10
- 239000012634 fragment Substances 0.000 claims description 10
- 101100450694 Arabidopsis thaliana HFR1 gene Proteins 0.000 claims description 9
- 239000000556 agonist Substances 0.000 claims description 8
- 238000006243 chemical reaction Methods 0.000 claims description 8
- 208000026278 immune system disease Diseases 0.000 claims description 8
- 208000030159 metabolic disease Diseases 0.000 claims description 8
- 238000012360 testing method Methods 0.000 claims description 7
- 229940124597 therapeutic agent Drugs 0.000 claims description 7
- 102000004127 Cytokines Human genes 0.000 claims description 6
- 108090000695 Cytokines Proteins 0.000 claims description 6
- 206010028980 Neoplasm Diseases 0.000 claims description 6
- 210000003527 eukaryotic cell Anatomy 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 6
- 210000004102 animal cell Anatomy 0.000 claims description 5
- 239000003937 drug carrier Substances 0.000 claims description 5
- 210000001236 prokaryotic cell Anatomy 0.000 claims description 5
- 241000588724 Escherichia coli Species 0.000 claims description 4
- 239000002260 anti-inflammatory agent Substances 0.000 claims description 4
- 229940121363 anti-inflammatory agent Drugs 0.000 claims description 4
- 239000002246 antineoplastic agent Substances 0.000 claims description 4
- 239000000872 buffer Substances 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 4
- 229940127089 cytotoxic agent Drugs 0.000 claims description 4
- 239000002955 immunomodulating agent Substances 0.000 claims description 4
- 229940121354 immunomodulator Drugs 0.000 claims description 4
- 230000002584 immunomodulator Effects 0.000 claims description 4
- 210000004962 mammalian cell Anatomy 0.000 claims description 4
- 230000005298 paramagnetic effect Effects 0.000 claims description 4
- 239000003053 toxin Substances 0.000 claims description 4
- 231100000765 toxin Toxicity 0.000 claims description 4
- 241000271566 Aves Species 0.000 claims description 3
- 241000238631 Hexapoda Species 0.000 claims description 3
- 102000018071 Immunoglobulin Fc Fragments Human genes 0.000 claims description 3
- 108010091135 Immunoglobulin Fc Fragments Proteins 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims description 3
- 208000016097 disease of metabolism Diseases 0.000 claims description 3
- 208000035475 disorder Diseases 0.000 claims description 3
- 239000007850 fluorescent dye Substances 0.000 claims description 3
- 230000002538 fungal effect Effects 0.000 claims description 3
- 239000005556 hormone Substances 0.000 claims description 3
- 229940088597 hormone Drugs 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims description 2
- 108010076504 Protein Sorting Signals Proteins 0.000 claims description 2
- 239000000032 diagnostic agent Substances 0.000 claims description 2
- 229940039227 diagnostic agent Drugs 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 230000003439 radiotherapeutic effect Effects 0.000 claims description 2
- 150000003839 salts Chemical class 0.000 claims description 2
- 238000013319 spin trapping Methods 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims 1
- 230000001225 therapeutic effect Effects 0.000 abstract description 6
- 238000002474 experimental method Methods 0.000 abstract description 2
- 241000700159 Rattus Species 0.000 description 28
- 239000008055 phosphate buffer solution Substances 0.000 description 16
- 239000002609 medium Substances 0.000 description 13
- 210000000988 bone and bone Anatomy 0.000 description 12
- 230000000694 effects Effects 0.000 description 10
- 210000004408 hybridoma Anatomy 0.000 description 10
- 230000003053 immunization Effects 0.000 description 10
- 238000002649 immunization Methods 0.000 description 10
- 102000004169 proteins and genes Human genes 0.000 description 10
- 108090000623 proteins and genes Proteins 0.000 description 10
- 241000699670 Mus sp. Species 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 9
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- 239000006285 cell suspension Substances 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- 239000000047 product Substances 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 239000007788 liquid Substances 0.000 description 6
- 210000000952 spleen Anatomy 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 5
- 238000002965 ELISA Methods 0.000 description 5
- 108050003558 Interleukin-17 Proteins 0.000 description 5
- 102000013691 Interleukin-17 Human genes 0.000 description 5
- 108090001005 Interleukin-6 Proteins 0.000 description 5
- 102000004889 Interleukin-6 Human genes 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 239000000427 antigen Substances 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 102000036639 antigens Human genes 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 230000010076 replication Effects 0.000 description 5
- 210000004989 spleen cell Anatomy 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- 210000005222 synovial tissue Anatomy 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 4
- 102000002689 Toll-like receptor Human genes 0.000 description 4
- 108020000411 Toll-like receptor Proteins 0.000 description 4
- 102100040247 Tumor necrosis factor Human genes 0.000 description 4
- 210000001015 abdomen Anatomy 0.000 description 4
- 239000002671 adjuvant Substances 0.000 description 4
- 210000001264 anterior cruciate ligament Anatomy 0.000 description 4
- 206010003246 arthritis Diseases 0.000 description 4
- 230000037182 bone density Effects 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 230000002757 inflammatory effect Effects 0.000 description 4
- 239000007928 intraperitoneal injection Substances 0.000 description 4
- 150000002500 ions Chemical class 0.000 description 4
- 210000000629 knee joint Anatomy 0.000 description 4
- 108020004999 messenger RNA Proteins 0.000 description 4
- 238000010603 microCT Methods 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 210000004417 patella Anatomy 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 238000005070 sampling Methods 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 4
- 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 3
- 238000013257 ACL transection animal model Methods 0.000 description 3
- 238000011725 BALB/c mouse Methods 0.000 description 3
- 102000003777 Interleukin-1 beta Human genes 0.000 description 3
- 108090000193 Interleukin-1 beta Proteins 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- -1 REG Chemical compound 0.000 description 3
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 3
- 241000607720 Serratia Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 3
- 238000010171 animal model Methods 0.000 description 3
- 230000000903 blocking effect Effects 0.000 description 3
- 238000010367 cloning Methods 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 239000003995 emulsifying agent Substances 0.000 description 3
- 239000013604 expression vector Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 3
- 230000028993 immune response Effects 0.000 description 3
- 210000000987 immune system Anatomy 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 239000000314 lubricant Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 239000000546 pharmaceutical excipient Substances 0.000 description 3
- 239000003755 preservative agent Substances 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 239000004017 serum-free culture medium Substances 0.000 description 3
- 210000001541 thymus gland Anatomy 0.000 description 3
- 108700012359 toxins Proteins 0.000 description 3
- VGIRNWJSIRVFRT-UHFFFAOYSA-N 2',7'-difluorofluorescein Chemical compound OC(=O)C1=CC=CC=C1C1=C2C=C(F)C(=O)C=C2OC2=CC(O)=C(F)C=C21 VGIRNWJSIRVFRT-UHFFFAOYSA-N 0.000 description 2
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 108010006654 Bleomycin Proteins 0.000 description 2
- COVZYZSDYWQREU-UHFFFAOYSA-N Busulfan Chemical compound CS(=O)(=O)OCCCCOS(C)(=O)=O COVZYZSDYWQREU-UHFFFAOYSA-N 0.000 description 2
- 102000000503 Collagen Type II Human genes 0.000 description 2
- 108010041390 Collagen Type II Proteins 0.000 description 2
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241000588914 Enterobacter Species 0.000 description 2
- 241000588921 Enterobacteriaceae Species 0.000 description 2
- 241000588698 Erwinia Species 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 2
- 101000669402 Homo sapiens Toll-like receptor 7 Proteins 0.000 description 2
- 241000588748 Klebsiella Species 0.000 description 2
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 2
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 2
- 241000588769 Proteus <enterobacteria> Species 0.000 description 2
- 241000589516 Pseudomonas Species 0.000 description 2
- 241000700157 Rattus norvegicus Species 0.000 description 2
- 241000607142 Salmonella Species 0.000 description 2
- 241000607768 Shigella Species 0.000 description 2
- 241000187747 Streptomyces Species 0.000 description 2
- NKANXQFJJICGDU-QPLCGJKRSA-N Tamoxifen Chemical compound C=1C=CC=CC=1C(/CC)=C(C=1C=CC(OCCN(C)C)=CC=1)/C1=CC=CC=C1 NKANXQFJJICGDU-QPLCGJKRSA-N 0.000 description 2
- 102100039390 Toll-like receptor 7 Human genes 0.000 description 2
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- RJURFGZVJUQBHK-UHFFFAOYSA-N actinomycin D Natural products CC1OC(=O)C(C(C)C)N(C)C(=O)CN(C)C(=O)C2CCCN2C(=O)C(C(C)C)NC(=O)C1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)NC4C(=O)NC(C(N5CCCC5C(=O)N(C)CC(=O)N(C)C(C(C)C)C(=O)OC4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-UHFFFAOYSA-N 0.000 description 2
- 210000001188 articular cartilage Anatomy 0.000 description 2
- 230000002146 bilateral effect Effects 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 229960001561 bleomycin Drugs 0.000 description 2
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCC[S+](C)C)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C OYVAGSVQBOHSSS-UAPAGMARSA-O 0.000 description 2
- 229960002092 busulfan Drugs 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 238000000423 cell based assay Methods 0.000 description 2
- 239000000460 chlorine Substances 0.000 description 2
- 239000011651 chromium Substances 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 229910017052 cobalt Inorganic materials 0.000 description 2
- 239000010941 cobalt Substances 0.000 description 2
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 229960000975 daunorubicin Drugs 0.000 description 2
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
- 229960004679 doxorubicin Drugs 0.000 description 2
- 238000004945 emulsification Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 238000012165 high-throughput sequencing Methods 0.000 description 2
- 210000002865 immune cell Anatomy 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 208000027866 inflammatory disease Diseases 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 239000007972 injectable composition Substances 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- SGDBTWWWUNNDEQ-LBPRGKRZSA-N melphalan Chemical compound OC(=O)[C@@H](N)CC1=CC=C(N(CCCl)CCCl)C=C1 SGDBTWWWUNNDEQ-LBPRGKRZSA-N 0.000 description 2
- 229960001924 melphalan Drugs 0.000 description 2
- 229920000609 methyl cellulose Polymers 0.000 description 2
- 239000001923 methylcellulose Substances 0.000 description 2
- 235000010981 methylcellulose Nutrition 0.000 description 2
- PXHVJJICTQNCMI-UHFFFAOYSA-N nickel Substances [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 2
- 230000036963 noncompetitive effect Effects 0.000 description 2
- 238000009806 oophorectomy Methods 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 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 2
- OARRHUQTFTUEOS-UHFFFAOYSA-N safranin Chemical compound [Cl-].C=12C=C(N)C(C)=CC2=NC2=CC(C)=C(N)C=C2[N+]=1C1=CC=CC=C1 OARRHUQTFTUEOS-UHFFFAOYSA-N 0.000 description 2
- 239000011669 selenium Substances 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 230000019491 signal transduction Effects 0.000 description 2
- 230000011664 signaling Effects 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- ABZLKHKQJHEPAX-UHFFFAOYSA-N tetramethylrhodamine Chemical compound C=12C=CC(N(C)C)=CC2=[O+]C2=CC(N(C)C)=CC=C2C=1C1=CC=CC=C1C([O-])=O ABZLKHKQJHEPAX-UHFFFAOYSA-N 0.000 description 2
- AUHDWARTFSKSAC-HEIFUQTGSA-N (2S,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)-2-(6-oxo-1H-purin-9-yl)oxolane-2-carboxylic acid Chemical compound [C@]1([C@H](O)[C@H](O)[C@@H](CO)O1)(N1C=NC=2C(O)=NC=NC12)C(=O)O AUHDWARTFSKSAC-HEIFUQTGSA-N 0.000 description 1
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 1
- HDCUSMZVZWLDRZ-UHFFFAOYSA-N 1-benzyl-2-methylhydrazine Chemical compound CNNCC1=CC=CC=C1 HDCUSMZVZWLDRZ-UHFFFAOYSA-N 0.000 description 1
- HANWHVWXFQSQGJ-UHFFFAOYSA-N 1-tetradecoxytetradecane Chemical compound CCCCCCCCCCCCCCOCCCCCCCCCCCCCC HANWHVWXFQSQGJ-UHFFFAOYSA-N 0.000 description 1
- PNDPGZBMCMUPRI-HVTJNCQCSA-N 10043-66-0 Chemical compound [131I][131I] PNDPGZBMCMUPRI-HVTJNCQCSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- IVLXQGJVBGMLRR-UHFFFAOYSA-N 2-aminoacetic acid;hydron;chloride Chemical compound Cl.NCC(O)=O IVLXQGJVBGMLRR-UHFFFAOYSA-N 0.000 description 1
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 1
- IDLISIVVYLGCKO-UHFFFAOYSA-N 6-carboxy-4',5'-dichloro-2',7'-dimethoxyfluorescein Chemical compound O1C(=O)C2=CC=C(C(O)=O)C=C2C21C1=CC(OC)=C(O)C(Cl)=C1OC1=C2C=C(OC)C(O)=C1Cl IDLISIVVYLGCKO-UHFFFAOYSA-N 0.000 description 1
- BZTDTCNHAFUJOG-UHFFFAOYSA-N 6-carboxyfluorescein Chemical compound C12=CC=C(O)C=C2OC2=CC(O)=CC=C2C11OC(=O)C2=CC=C(C(=O)O)C=C21 BZTDTCNHAFUJOG-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 241000607534 Aeromonas Species 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 101710165425 Alpha-enolase Proteins 0.000 description 1
- 206010002556 Ankylosing Spondylitis Diseases 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 241000194108 Bacillus licheniformis Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 108030001720 Bontoxilysin Proteins 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 101100244725 Caenorhabditis elegans pef-1 gene Proteins 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 108010049048 Cholera Toxin Proteins 0.000 description 1
- 102000009016 Cholera Toxin Human genes 0.000 description 1
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- 208000011231 Crohn disease Diseases 0.000 description 1
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 description 1
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 1
- 102000010831 Cytoskeletal Proteins Human genes 0.000 description 1
- 108010037414 Cytoskeletal Proteins Proteins 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 102000003844 DNA helicases Human genes 0.000 description 1
- 108090000133 DNA helicases Proteins 0.000 description 1
- 108010092160 Dactinomycin Proteins 0.000 description 1
- 241000702421 Dependoparvovirus Species 0.000 description 1
- 229910052692 Dysprosium Inorganic materials 0.000 description 1
- 206010014733 Endometrial cancer Diseases 0.000 description 1
- 206010014759 Endometrial neoplasm Diseases 0.000 description 1
- 101710184673 Enolase 1 Proteins 0.000 description 1
- 229910052691 Erbium Inorganic materials 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- 241000588722 Escherichia Species 0.000 description 1
- 241000701959 Escherichia virus Lambda Species 0.000 description 1
- 241001524679 Escherichia virus M13 Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 229910052693 Europium Inorganic materials 0.000 description 1
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 1
- 108010040721 Flagellin Proteins 0.000 description 1
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 1
- 229910052688 Gadolinium Inorganic materials 0.000 description 1
- 208000007882 Gastritis Diseases 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 201000005569 Gout Diseases 0.000 description 1
- 208000009329 Graft vs Host Disease Diseases 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
- 208000001204 Hashimoto Disease Diseases 0.000 description 1
- 208000030836 Hashimoto thyroiditis Diseases 0.000 description 1
- 229910052689 Holmium Inorganic materials 0.000 description 1
- 101000800483 Homo sapiens Toll-like receptor 8 Proteins 0.000 description 1
- 101000611183 Homo sapiens Tumor necrosis factor Proteins 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 208000022559 Inflammatory bowel disease Diseases 0.000 description 1
- GRSZFWQUAKGDAV-UHFFFAOYSA-N Inosinic acid Natural products OC1C(O)C(COP(O)(O)=O)OC1N1C(NC=NC2=O)=C2N=C1 GRSZFWQUAKGDAV-UHFFFAOYSA-N 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 108090000174 Interleukin-10 Proteins 0.000 description 1
- 108090000177 Interleukin-11 Proteins 0.000 description 1
- 108010065805 Interleukin-12 Proteins 0.000 description 1
- 108090000176 Interleukin-13 Proteins 0.000 description 1
- 108090000172 Interleukin-15 Proteins 0.000 description 1
- 101800003050 Interleukin-16 Proteins 0.000 description 1
- 108090000171 Interleukin-18 Proteins 0.000 description 1
- 108050009288 Interleukin-19 Proteins 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- 108010065637 Interleukin-23 Proteins 0.000 description 1
- 108010002386 Interleukin-3 Proteins 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 108010002616 Interleukin-5 Proteins 0.000 description 1
- 108010002586 Interleukin-7 Proteins 0.000 description 1
- 108090001007 Interleukin-8 Proteins 0.000 description 1
- 108010002335 Interleukin-9 Proteins 0.000 description 1
- ZCYVEMRRCGMTRW-AHCXROLUSA-N Iodine-123 Chemical compound [123I] ZCYVEMRRCGMTRW-AHCXROLUSA-N 0.000 description 1
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 1
- YQEZLKZALYSWHR-UHFFFAOYSA-N Ketamine Chemical compound C=1C=CC=C(Cl)C=1C1(NC)CCCCC1=O YQEZLKZALYSWHR-UHFFFAOYSA-N 0.000 description 1
- 208000008839 Kidney Neoplasms Diseases 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 244000178870 Lavandula angustifolia Species 0.000 description 1
- 235000010663 Lavandula angustifolia Nutrition 0.000 description 1
- 241000713666 Lentivirus Species 0.000 description 1
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 1
- 240000007472 Leucaena leucocephala Species 0.000 description 1
- 102000043136 MAP kinase family Human genes 0.000 description 1
- 108091054455 MAP kinase family Proteins 0.000 description 1
- 208000002720 Malnutrition Diseases 0.000 description 1
- WAEMQWOKJMHJLA-UHFFFAOYSA-N Manganese(2+) Chemical compound [Mn+2] WAEMQWOKJMHJLA-UHFFFAOYSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 229930192392 Mitomycin Natural products 0.000 description 1
- 239000004909 Moisturizer Substances 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 1
- VEQPNABPJHWNSG-UHFFFAOYSA-N Nickel(2+) Chemical compound [Ni+2] VEQPNABPJHWNSG-UHFFFAOYSA-N 0.000 description 1
- AWZJFZMWSUBJAJ-UHFFFAOYSA-N OG-514 dye Chemical compound OC(=O)CSC1=C(F)C(F)=C(C(O)=O)C(C2=C3C=C(F)C(=O)C=C3OC3=CC(O)=C(F)C=C32)=C1F AWZJFZMWSUBJAJ-UHFFFAOYSA-N 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 241001631646 Papillomaviridae Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 108010081690 Pertussis Toxin Proteins 0.000 description 1
- 108010022181 Phosphopyruvate Hydratase Proteins 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 208000003286 Protein-Energy Malnutrition Diseases 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 238000010802 RNA extraction kit Methods 0.000 description 1
- 206010038389 Renal cancer Diseases 0.000 description 1
- 108700008625 Reporter Genes Proteins 0.000 description 1
- 108010039491 Ricin Proteins 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- 241000607715 Serratia marcescens Species 0.000 description 1
- 241000700584 Simplexvirus Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- GKLVYJBZJHMRIY-OUBTZVSYSA-N Technetium-99 Chemical compound [99Tc] GKLVYJBZJHMRIY-OUBTZVSYSA-N 0.000 description 1
- 229910052771 Terbium Inorganic materials 0.000 description 1
- 108010060818 Toll-Like Receptor 9 Proteins 0.000 description 1
- 102100033110 Toll-like receptor 8 Human genes 0.000 description 1
- 102100033117 Toll-like receptor 9 Human genes 0.000 description 1
- GYDJEQRTZSCIOI-UHFFFAOYSA-N Tranexamic acid Chemical compound NCC1CCC(C(O)=O)CC1 GYDJEQRTZSCIOI-UHFFFAOYSA-N 0.000 description 1
- JXLYSJRDGCGARV-WWYNWVTFSA-N Vinblastine Natural products O=C(O[C@H]1[C@](O)(C(=O)OC)[C@@H]2N(C)c3c(cc(c(OC)c3)[C@]3(C(=O)OC)c4[nH]c5c(c4CCN4C[C@](O)(CC)C[C@H](C3)C4)cccc5)[C@@]32[C@H]2[C@@]1(CC)C=CCN2CC3)C JXLYSJRDGCGARV-WWYNWVTFSA-N 0.000 description 1
- 206010047626 Vitamin D Deficiency Diseases 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 229910052769 Ytterbium Inorganic materials 0.000 description 1
- VWQVUPCCIRVNHF-OUBTZVSYSA-N Yttrium-90 Chemical compound [90Y] VWQVUPCCIRVNHF-OUBTZVSYSA-N 0.000 description 1
- RJURFGZVJUQBHK-IIXSONLDSA-N actinomycin D Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)N[C@@H]4C(=O)N[C@@H](C(N5CCC[C@H]5C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-IIXSONLDSA-N 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 229940035676 analgesics Drugs 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 239000000730 antalgic agent Substances 0.000 description 1
- 238000011091 antibody purification Methods 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- RYXHOMYVWAEKHL-OUBTZVSYSA-N astatine-211 Chemical compound [211At] RYXHOMYVWAEKHL-OUBTZVSYSA-N 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- VSRXQHXAPYXROS-UHFFFAOYSA-N azanide;cyclobutane-1,1-dicarboxylic acid;platinum(2+) Chemical compound [NH2-].[NH2-].[Pt+2].OC(=O)C1(C(O)=O)CCC1 VSRXQHXAPYXROS-UHFFFAOYSA-N 0.000 description 1
- 238000010009 beating Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 229940053031 botulinum toxin Drugs 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000000378 calcium silicate Substances 0.000 description 1
- 229910052918 calcium silicate Inorganic materials 0.000 description 1
- 235000012241 calcium silicate Nutrition 0.000 description 1
- OYACROKNLOSFPA-UHFFFAOYSA-N calcium;dioxido(oxo)silane Chemical compound [Ca+2].[O-][Si]([O-])=O OYACROKNLOSFPA-UHFFFAOYSA-N 0.000 description 1
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 229960004562 carboplatin Drugs 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- CZPLANDPABRVHX-UHFFFAOYSA-N cascade blue Chemical compound C=1C2=CC=CC=C2C(NCC)=CC=1C(C=1C=CC(=CC=1)N(CC)CC)=C1C=CC(=[N+](CC)CC)C=C1 CZPLANDPABRVHX-UHFFFAOYSA-N 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
- 229960002327 chloral hydrate Drugs 0.000 description 1
- 229960004630 chlorambucil Drugs 0.000 description 1
- JCKYGMPEJWAADB-UHFFFAOYSA-N chlorambucil Chemical compound OC(=O)CCCC1=CC=C(N(CCCl)CCCl)C=C1 JCKYGMPEJWAADB-UHFFFAOYSA-N 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 229910052804 chromium Inorganic materials 0.000 description 1
- BFGKITSFLPAWGI-UHFFFAOYSA-N chromium(3+) Chemical compound [Cr+3] BFGKITSFLPAWGI-UHFFFAOYSA-N 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- 239000013599 cloning vector Substances 0.000 description 1
- XLJKHNWPARRRJB-UHFFFAOYSA-N cobalt(2+) Chemical compound [Co+2] XLJKHNWPARRRJB-UHFFFAOYSA-N 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- RYGMFSIKBFXOCR-AKLPVKDBSA-N copper-67 Chemical compound [67Cu] RYGMFSIKBFXOCR-AKLPVKDBSA-N 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 229960000640 dactinomycin Drugs 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 108010017271 denileukin diftitox Proteins 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- HRLIOXLXPOHXTA-NSHDSACASA-N dexmedetomidine Chemical compound C1([C@@H](C)C=2C(=C(C)C=CC=2)C)=CN=C[N]1 HRLIOXLXPOHXTA-NSHDSACASA-N 0.000 description 1
- 229960004253 dexmedetomidine Drugs 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- QYHNIMDZIYANJH-UHFFFAOYSA-N diindium Chemical compound [In]#[In] QYHNIMDZIYANJH-UHFFFAOYSA-N 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- KBQHZAAAGSGFKK-UHFFFAOYSA-N dysprosium atom Chemical compound [Dy] KBQHZAAAGSGFKK-UHFFFAOYSA-N 0.000 description 1
- 230000002876 effect on osteoporosis Effects 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- UYAHIZSMUZPPFV-UHFFFAOYSA-N erbium Chemical compound [Er] UYAHIZSMUZPPFV-UHFFFAOYSA-N 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- 102000015694 estrogen receptors Human genes 0.000 description 1
- 108010038795 estrogen receptors Proteins 0.000 description 1
- VJJPUSNTGOMMGY-MRVIYFEKSA-N etoposide Chemical compound COC1=C(O)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@@H](O[C@H]3[C@@H]([C@@H](O)[C@@H]4O[C@H](C)OC[C@H]4O3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 VJJPUSNTGOMMGY-MRVIYFEKSA-N 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 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 1
- 229960002949 fluorouracil Drugs 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- UIWYJDYFSGRHKR-UHFFFAOYSA-N gadolinium atom Chemical compound [Gd] UIWYJDYFSGRHKR-UHFFFAOYSA-N 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 229960005277 gemcitabine Drugs 0.000 description 1
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000034659 glycolysis Effects 0.000 description 1
- 208000024908 graft versus host disease Diseases 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 201000010536 head and neck cancer Diseases 0.000 description 1
- 208000014829 head and neck neoplasm Diseases 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- KJZYNXUDTRRSPN-UHFFFAOYSA-N holmium atom Chemical compound [Ho] KJZYNXUDTRRSPN-UHFFFAOYSA-N 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- HOMGKSMUEGBAAB-UHFFFAOYSA-N ifosfamide Chemical compound ClCCNP1(=O)OCCCN1CCCl HOMGKSMUEGBAAB-UHFFFAOYSA-N 0.000 description 1
- 229960001101 ifosfamide Drugs 0.000 description 1
- 229960002751 imiquimod Drugs 0.000 description 1
- DOUYETYNHWVLEO-UHFFFAOYSA-N imiquimod Chemical compound C1=CC=CC2=C3N(CC(C)C)C=NC3=C(N)N=C21 DOUYETYNHWVLEO-UHFFFAOYSA-N 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 229960000598 infliximab Drugs 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000004245 inosinic acid Substances 0.000 description 1
- 229940028843 inosinic acid Drugs 0.000 description 1
- 235000013902 inosinic acid Nutrition 0.000 description 1
- 102000006495 integrins Human genes 0.000 description 1
- 108010044426 integrins Proteins 0.000 description 1
- 108090000681 interleukin 20 Proteins 0.000 description 1
- 108010074108 interleukin-21 Proteins 0.000 description 1
- 108010074109 interleukin-22 Proteins 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- XMBWDFGMSWQBCA-YPZZEJLDSA-N iodane Chemical compound [125IH] XMBWDFGMSWQBCA-YPZZEJLDSA-N 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 229960002725 isoflurane Drugs 0.000 description 1
- 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 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 229960003299 ketamine Drugs 0.000 description 1
- 201000010982 kidney cancer Diseases 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000001102 lavandula vera Substances 0.000 description 1
- 235000018219 lavender Nutrition 0.000 description 1
- GZQKNULLWNGMCW-PWQABINMSA-N lipid A (E. coli) Chemical compound O1[C@H](CO)[C@@H](OP(O)(O)=O)[C@H](OC(=O)C[C@@H](CCCCCCCCCCC)OC(=O)CCCCCCCCCCCCC)[C@@H](NC(=O)C[C@@H](CCCCCCCCCCC)OC(=O)CCCCCCCCCCC)[C@@H]1OC[C@@H]1[C@@H](O)[C@H](OC(=O)C[C@H](O)CCCCCCCCCCC)[C@@H](NC(=O)C[C@H](O)CCCCCCCCCCC)[C@@H](OP(O)(O)=O)O1 GZQKNULLWNGMCW-PWQABINMSA-N 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 230000007762 localization of cell Effects 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 239000000845 maltitol Substances 0.000 description 1
- 235000010449 maltitol Nutrition 0.000 description 1
- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 description 1
- 229940035436 maltitol Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- HAWPXGHAZFHHAD-UHFFFAOYSA-N mechlorethamine Chemical class ClCCN(C)CCCl HAWPXGHAZFHHAD-UHFFFAOYSA-N 0.000 description 1
- 229960004961 mechlorethamine Drugs 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- CFCUWKMKBJTWLW-BKHRDMLASA-N mithramycin Chemical compound O([C@@H]1C[C@@H](O[C@H](C)[C@H]1O)OC=1C=C2C=C3C[C@H]([C@@H](C(=O)C3=C(O)C2=C(O)C=1C)O[C@@H]1O[C@H](C)[C@@H](O)[C@H](O[C@@H]2O[C@H](C)[C@H](O)[C@H](O[C@@H]3O[C@H](C)[C@@H](O)[C@@](C)(O)C3)C2)C1)[C@H](OC)C(=O)[C@@H](O)[C@@H](C)O)[C@H]1C[C@@H](O)[C@H](O)[C@@H](C)O1 CFCUWKMKBJTWLW-BKHRDMLASA-N 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- 108091005573 modified proteins Proteins 0.000 description 1
- 102000035118 modified proteins Human genes 0.000 description 1
- 230000001333 moisturizer Effects 0.000 description 1
- 229940035032 monophosphoryl lipid a Drugs 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 201000006417 multiple sclerosis Diseases 0.000 description 1
- QEFYFXOXNSNQGX-UHFFFAOYSA-N neodymium atom Chemical compound [Nd] QEFYFXOXNSNQGX-UHFFFAOYSA-N 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- OSTGTTZJOCZWJG-UHFFFAOYSA-N nitrosourea Chemical compound NC(=O)N=NO OSTGTTZJOCZWJG-UHFFFAOYSA-N 0.000 description 1
- 239000000041 non-steroidal anti-inflammatory agent Substances 0.000 description 1
- 229940021182 non-steroidal anti-inflammatory drug Drugs 0.000 description 1
- 229940046166 oligodeoxynucleotide Drugs 0.000 description 1
- 229940100027 ontak Drugs 0.000 description 1
- 230000001009 osteoporotic effect Effects 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- VYNDHICBIRRPFP-UHFFFAOYSA-N pacific blue Chemical compound FC1=C(O)C(F)=C2OC(=O)C(C(=O)O)=CC2=C1 VYNDHICBIRRPFP-UHFFFAOYSA-N 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 210000004303 peritoneum Anatomy 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229960003171 plicamycin Drugs 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 235000020826 protein-energy malnutrition Nutrition 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- GZUITABIAKMVPG-UHFFFAOYSA-N raloxifene Chemical compound C1=CC(O)=CC=C1C1=C(C(=O)C=2C=CC(OCCN3CCCCC3)=CC=2)C2=CC=C(O)C=C2S1 GZUITABIAKMVPG-UHFFFAOYSA-N 0.000 description 1
- 229960004622 raloxifene Drugs 0.000 description 1
- ZAHRKKWIAAJSAO-UHFFFAOYSA-N rapamycin Natural products COCC(O)C(=C/C(C)C(=O)CC(OC(=O)C1CCCCN1C(=O)C(=O)C2(O)OC(CC(OC)C(=CC=CC=CC(C)CC(C)C(=O)C)C)CCC2C)C(C)CC3CCC(O)C(C3)OC)C ZAHRKKWIAAJSAO-UHFFFAOYSA-N 0.000 description 1
- 238000011552 rat model Methods 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000002271 resection Methods 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 229910052702 rhenium Inorganic materials 0.000 description 1
- WUAPFZMCVAUBPE-UHFFFAOYSA-N rhenium atom Chemical compound [Re] WUAPFZMCVAUBPE-UHFFFAOYSA-N 0.000 description 1
- WUAPFZMCVAUBPE-IGMARMGPSA-N rhenium-186 Chemical compound [186Re] WUAPFZMCVAUBPE-IGMARMGPSA-N 0.000 description 1
- DOSGOCSVHPUUIA-UHFFFAOYSA-N samarium(3+) Chemical compound [Sm+3] DOSGOCSVHPUUIA-UHFFFAOYSA-N 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 229960002930 sirolimus Drugs 0.000 description 1
- QFJCIRLUMZQUOT-HPLJOQBZSA-N sirolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1C[C@@H](C)[C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@](O)(O2)[C@H](C)CC[C@H]2C[C@H](OC)/C(C)=C/C=C/C=C/[C@@H](C)C[C@@H](C)C(=O)[C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(=O)C1 QFJCIRLUMZQUOT-HPLJOQBZSA-N 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 210000005065 subchondral bone plate Anatomy 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 201000000596 systemic lupus erythematosus Diseases 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 229960001603 tamoxifen Drugs 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- GZCRRIHWUXGPOV-UHFFFAOYSA-N terbium atom Chemical compound [Tb] GZCRRIHWUXGPOV-UHFFFAOYSA-N 0.000 description 1
- MPLHNVLQVRSVEE-UHFFFAOYSA-N texas red Chemical compound [O-]S(=O)(=O)C1=CC(S(Cl)(=O)=O)=CC=C1C(C1=CC=2CCCN3CCCC(C=23)=C1O1)=C2C1=C(CCC1)C3=[N+]1CCCC3=C2 MPLHNVLQVRSVEE-UHFFFAOYSA-N 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 210000002303 tibia Anatomy 0.000 description 1
- 238000011200 topical administration Methods 0.000 description 1
- 230000005026 transcription initiation Effects 0.000 description 1
- 239000003558 transferase inhibitor Substances 0.000 description 1
- 230000005945 translocation Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 210000002993 trophoblast Anatomy 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 241001529453 unidentified herpesvirus Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- 229910052720 vanadium Inorganic materials 0.000 description 1
- LEONUFNNVUYDNQ-UHFFFAOYSA-N vanadium atom Chemical compound [V] LEONUFNNVUYDNQ-UHFFFAOYSA-N 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 229960003048 vinblastine Drugs 0.000 description 1
- JXLYSJRDGCGARV-XQKSVPLYSA-N vincaleukoblastine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 JXLYSJRDGCGARV-XQKSVPLYSA-N 0.000 description 1
- 229960004528 vincristine Drugs 0.000 description 1
- OGWKCGZFUXNPDA-XQKSVPLYSA-N vincristine Chemical compound C([N@]1C[C@@H](C[C@]2(C(=O)OC)C=3C(=CC4=C([C@]56[C@H]([C@@]([C@H](OC(C)=O)[C@]7(CC)C=CCN([C@H]67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)C[C@@](C1)(O)CC)CC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-XQKSVPLYSA-N 0.000 description 1
- OGWKCGZFUXNPDA-UHFFFAOYSA-N vincristine Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(OC(C)=O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-UHFFFAOYSA-N 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 235000012431 wafers Nutrition 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- NAWDYIZEMPQZHO-UHFFFAOYSA-N ytterbium Chemical compound [Yb] NAWDYIZEMPQZHO-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/40—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- 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
- A61K47/68—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 antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6835—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 antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
- A61K47/6871—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 antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting an enzyme
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding 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
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/565—Complementarity determining region [CDR]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/567—Framework region [FR]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/988—Lyases (4.), e.g. aldolases, heparinase, enolases, fumarase
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Physical Education & Sports Medicine (AREA)
- Rheumatology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Epidemiology (AREA)
- Analytical Chemistry (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Pain & Pain Management (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明公开了一种ENO1抗体及其应用,所述抗体包括分别如SEQ ID NO:1、2、3所示的氨基酸序列的重链可变区HCDR1、HCDR2、HCDR3;以及分别如SEQ ID NO:9、10、11所示的氨基酸序列的轻链可变区LCDR1、LCDR2、LCDR3。本发明同时公开了编码所述抗体的核酸分子、含有所述核酸分子的载体和宿主细胞,经实验证明本发明的抗体对骨关节炎、骨质疏松及类风湿性关节炎具有良好的治疗效果。
Description
技术领域
本发明属于生物医药领域,具体涉及一种ENO1抗体及其应用。
背景技术
ENO1(α-烯醇化酶,烯醇化酶-1)是一种多功能蛋白,它的功能与细胞定位有关。位于细胞表面的ENO1是蛋白复合物(包括uPA受体、整合素和细胞骨架蛋白)的一部分,可促进肿瘤细胞的侵袭和转移;位于细胞质可参与糖酵解过程;此外,ENO1还可表达在细胞核中,与c-myc启动子结合,负向调控c-myc的表达,发挥抗肿瘤作用。
研究表明,ENO1的移位受到MAP激酶信号传导通路的调节,这表明细胞表面上ENO1表达的增加可能在炎症疾病中起到重要作用,已在不同自身免疫和炎性疾病中发现抗ENO1的自身抗体。抗体以其独特的生物学活性在疾病治疗中得到广泛应用。因此,研发新的抗ENO1的抗体对于实现ENO1相关疾病的检测和治疗具有重要的作用。
发明内容
为弥补现有技术的不足,本发明提供了一种抗ENO1的单克隆抗体及其在治疗疾病中的应用。
为实现上述目的,本发明采用如下技术方案:
本发明的第一方面提供了一种抗ENO1的单克隆抗体,所述单克隆抗体包括具有与SEQ ID NO:1、2、3所示的氨基酸序列至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的重链可变区HCDR1、HCDR2、HCDR3,或其具有一个或多个保守氨基酸取代的任何变体;
以及具有与SEQ ID NO:9、10、11所示的氨基酸序列至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的轻链可变区LCDR1、LCDR2、LCDR3,或其具有一个或多个保守氨基酸取代的任何变体。
进一步,所述HCDR1、HCDR2、HCDR3的氨基酸序列分别如SEQ ID NO:1、2、3所示;
所述LCDR1、LCDR2、LCDR3的氨基酸序列分别如SEQ ID NO:9、10、11所示。
进一步,所述单克隆抗体还包括具有与SEQ ID NO:4、5、6、7所示的氨基酸序列至少90%、至少92%、至少93%、至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的重链可变区框架区HFR1、HFR2、HFR3、HFR4,或其具有一个或多个保守氨基酸取代的任何变体;
以及具有与SEQ ID NO:12、13、14、15所示的氨基酸序列至少90%、至少92%、至少93%、至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的轻链可变区框架区LFR1、LFR2、LFR3、LFR4,或其具有一个或多个保守氨基酸取代的任何变体。
进一步,所述重链可变区框架区HFR1、HFR2、HFR3、HFR4的氨基酸序列分别如SEQID NO:4、5、6、7所示;
所述轻链可变区框架区LFR1、LFR2、LFR3、LFR4的氨基酸序列分别如SEQ ID NO:12、13、14、15所示。
进一步,所述单克隆抗体重链可变区具有与SEQ ID NO:8所示的氨基酸序列至少70%、至少75%、至少80%、至少85%、至少90%、至少92%、至少93%、95%、至少96%、至少97%、至少98%、至少99%序列同一性的VH,或其具有一个或多个保守氨基酸取代的任何变体;
所述轻链可变区具有与SEQ ID NO:16所示的氨基酸序列至少70%、至少75%、至少80%、至少85%、至少90%、至少92%、至少93%、95%、至少96%、至少97%、至少98%、至少99%序列同一性的VL,或其具有一个或多个保守氨基酸取代的任何变体。
进一步,所述单克隆抗体的VH具有如SEQ ID NO:8所示的氨基酸序列,所述单克隆抗体的VL具有如SEQ ID NO:16所示的氨基酸序列。
进一步,所述单克隆抗体还包括抗体重链恒定区和/或抗体轻链恒定区的全部或者部分。
进一步,所述单克隆抗体的重链亚型为IgG2。
进一步,所述IgG2包括IgG2a、IgG2b。
进一步,所述单克隆抗体的重链亚型为IgG2a。
进一步,所述单克隆抗体的轻链亚型为κ。
本发明的第二方面提供了如下任一项所述的物质:
(1)核酸分子,所述核酸分子编码本发明第一方面所述的单克隆抗体或其功能片段;
(2)载体,所述载体包括(1)中所述的核酸分子;
(3)宿主细胞,所述宿主细胞包括(1)中所述的核酸分子或(2)中所述的载体;
(4)药物偶联物,所述药物偶联物包括本发明第一方面所述的单克隆抗体或其功能片段;
(5)一种检测ENO1的产品,所述产品包括本发明第一方面所述的单克隆抗体或其功能片段。
进一步,(1)中编码HCDR1、HCDR2、HCDR3的核酸分子具有与SEQ ID NO:17、18、19所示的核苷酸序列至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列;
编码LCDR1、LCDR2、LCDR3的核酸分子具有与SEQ ID NO:25、26、27所示的核苷酸序列至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列。
进一步,编码HCDR1、HCDR2、HCDR3的核酸分子的核苷酸序列分别如SEQ ID NO:17、18、19所示,编码LCDR1、LCDR2、LCDR3的核酸分子的核苷酸序列分别如SEQ ID NO:25、26、27所示。
进一步,编码HFR1、HFR2、HFR3、HFR4的核酸分子具有与SEQ ID NO:20、21、22、23所示的核苷酸序列至少90%、至少92%、至少93%、至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列;
编码LFR1、LFR2、LFR3、LFR4的核酸分子具有与SEQ ID NO:28、29、30、31所示的核苷酸序列至少90%、至少92%、至少93%、至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列。
进一步,编码HFR1、HFR2、HFR3、HFR4的核酸分子的核苷酸序列分别如SEQ ID NO:20、21、22、23所示,编码LFR1、LFR2、LFR3、LFR4的核酸分子的核苷酸序列分别如SEQ ID NO:28、29、30、31所示。
进一步,编码VH的核酸分子具有与SEQ ID NO:24所示的核苷酸序列至少70%、至少75%、至少80%、至少85%、至少90%、至少92%、至少93%、95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列;
编码VL的核酸分子具有与SEQ ID NO:32所示的核苷酸序列至少70%、至少75%、至少80%、至少85%、至少90%、至少92%、至少93%、95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列。
进一步,编码VH的核酸分子的核苷酸序列如SEQ ID NO:24所示,编码VL的核酸分子的核苷酸序列如SEQ ID NO:32所示。
进一步,(2)中所述的载体具有与抗体可操作的连接的信号肽。
进一步,所述载体还包括用于控制表达的元件。
进一步,所述载体还包括有助于其进入细胞的材料。
进一步,(3)中所述的宿主细胞包括原核细胞、真核细胞。
进一步,所述原核细胞包括大肠杆菌。
进一步,所述真核细胞包括原生生物细胞、动物细胞、植物细胞或真菌细胞。
进一步,所述动物细胞包括哺乳动物细胞、鸟类细胞、昆虫细胞。
进一步,(4)中所述的药物偶联物还包括可检测标记物、药物、毒素、细胞因子和/或酶。
进一步,(5)中所述的产品还包括用于执行抗原-抗体反应的试剂或用于检测反应的试剂。
进一步,用于执行抗原-抗体反应的试剂包括缓冲剂、盐。
进一步,所述产品包括试剂盒、试纸条。
进一步,所述试剂盒包括连接有信号产生化合物的抗体的缀合物。
本发明的第三方面提供了一种衍生物,所述衍生物包括在本发明第一方面所述的单克隆抗体或本发明第二方面所述的核酸分子上连接诊断试剂、治疗试剂。
进一步,所述诊断试剂包括荧光标记、放射性同位素、化学发光分子、顺磁性离子或自旋捕获试剂。
进一步,所述治疗试剂包括细胞因子、化学治疗剂、放射治疗剂、抗炎剂、激素、抗体Fc片段、TLR激动剂、含CpG的分子或免疫共刺激分子。
本发明的第四方面提供了如下任一项所述的方法:
(1)一种制备本发明第一方面所述的单克隆抗体的方法,所述方法包括:培养本发明第二方面所述的宿主细胞,回收单克隆抗体;
(2)一种检测样本中ENO1的方法,所述方法包括:使本发明第一方面所述的抗ENO1的单克隆抗体与待测样本接触,从而检测待测样本中ENO1的水平。
进一步,(1)中所述的方法还包括纯化所述的单克隆抗体。
本发明的第五方面提供了一种药物组合物,所述药物组合物包括本发明第一方面所述的单克隆抗体、本发明第二方面所述的物质或本发明第三方面所述的衍生物。
进一步,所述药物组合物还包括免疫调节剂。
进一步,所述药物组合物还包括药学上可接受的载体。
本发明的第六方面提供了如下任一项所述的应用:
(1)本发明第一方面所述的单克隆抗体、本发明第二方面所述的物质或本发明第三方面所述的衍生物在检测ENO1中的应用;
(2)本发明第一方面所述的单克隆抗体、本发明第二方面所述的物质或本发明第三方面所述的衍生物在制备诊断ENO1相关疾病的产品中的应用;
(3)本发明第一方面所述的单克隆抗体、本发明第二方面所述的物质、本发明第三方面所述的衍生物或本发明第五方面的药物组合物在制备预防和/或治疗ENO1相关疾病的药物中的应用。
进一步,所述ENO1相关疾病包括肿瘤、代谢性疾病和/或免疫疾病。
进一步,所述ENO1相关疾病选自代谢性疾病和/或免疫疾病。
进一步,所述免疫疾病包括骨关节炎、类风湿关节炎。
进一步,所述代谢性疾病选自骨质疏松。
本发明的优点和有益效果:
本发明提供了一种ENO1抗体及其应用,所述抗体对ENO1具有较高的亲和活性,对骨质疏松、骨关节炎、类风湿性关节炎等具有良好的治疗效果。
附图说明
图1是ENO1蛋白的表达图;
图2是ENO1蛋白洗脱条件图;
图3是抗ENO1单抗的蛋白纯度鉴定图;
图4是ENO1单抗的亚型图;
图5是23G5的KD值检测图;
图6是左侧胫骨平台软骨下内侧骨矢状图;
图7是小鼠骨关节炎参数图,其中,7A是骨密度图,7B是骨体积/组织体积分数图,7C是孔隙率图,7D是结构模型指数图,7E是骨小梁厚度图,7F是骨小梁数目图,7G是骨小梁分离图,7H是骨小梁模式因素图;
图8是HE和Safranin O染色图;
图9是大鼠的股骨干远端micro-CT图;
图10是大鼠骨质疏松参数图,其中,10A是骨体积/组织体积分数图,10B是骨小梁数目图,10C是骨小梁厚度图,10D是骨小梁分离度图,10E是骨密度图;
图11是大鼠关节炎评分图;
图12是大鼠滑膜组织中炎症因子mRNA相对水平图,其中,12A是TNF-αmRNA相对水平图,12B是IL-6mRNA相对水平图,12C是IL-1βmRNA相对水平图,12D是IL-17mRNA相对水平图。
具体实施方式
下文提供了本说明书中使用的一些术语的定义。除非另有说明,本文中使用的所有技术和科学用语通常具有和本发明所属领域的普通技术人员通常理解的意思相同的意思。
本发明提供了一种抗ENO1的单克隆抗体,所述单克隆抗体包括重链可变区HCDR1、HCDR2、HCDR3;和轻链可变区LCDR1、LCDR2、LCDR3。
在本发明中,单克隆抗体(monoclonal Ab,mAB,单抗)是指具有单一分子组成的抗体分子,获自一群基本相同的抗体。抗体包含两条重(H)链和两条轻(L)链。哺乳动物重链由可变区(VH)以及第一、第二、第三和任选存在的第四恒定区(分别为CH1、CH2、CH3、CH4)组成;哺乳动物轻链由可变区(VL)和恒定区组成。抗体呈Y形,其中Y的茎部由通过二硫键结合在一起的两条重链的第二和第三恒定区组成。Y的每个臂包括与单一轻链的可变区和恒定区结合的单一重链的可变区和第一恒定区。轻链和重链的可变区负责抗原结合。两条链中的可变区一般含有三个高度可变的环,称为互补决定区(complementaritydeterminingregion;CDR),轻链CDR包括LCDR1、LCDR2和LCDR3,重链CDR包括HCDR1、HCDR2、HCDR3。轻链和重链的可变区还包括框架区(framework region;FR),轻链FR包括LFR1、LFR2、LFR3和LFR4,重链FR包括HFR1、HFR2、HFR3和HFR4。重链和轻链的恒定区不参与抗原结合,但呈现出各种效应功能。基于抗体重链恒定区的氨基酸序列来对抗体分类。通常,抗体的种类由重链决定。例如哺乳动物的轻链分类为λ或κ,重链一共有五种,分别用希腊字母α、δ、ε、γ和μ来命名,相对应组成的抗体就称为IgA、IgD、IgE、IgG和IgM。其中,人的γ可以进一步细分为γ1、γ2、γ3、γ4等亚类,分别对应IgG1、IgG2、IgG3和IgG4四个亚型,小鼠γ可以进一步细分为γ1、γ2a、γ2b、γ3等亚类,分别对应IgG1、IgG2a、IgG2b和IgG3四个亚型。
本发明的抗体具有一个或多个保守氨基酸取代的任何变体,所述变体是指由于亲本抗体序列中一个或多个氨基酸残基的添加、缺失和/或取代而在氨基酸序列上与亲本抗ENO1抗体氨基酸序列不同并且保留亲本抗ENO1抗体的至少一种所期望的活性的分子。所期望的活性可以包括特异地结合抗原的能力、降低、抑制或中和动物中ENO1活性的能力和在基于细胞的测定中抑制ENO1介导的信号传导的能力。在一个实施例中,变体在亲本抗体的一个或多个高变区和/或构架区中包含一个或多个氨基酸取代。例如,变体可以在亲本抗体的一个或多个高变区和/或构架区中包含至少一个,或约一个至约十个,或约两个至约五个取代。变体保留了结合ENO1变体的能力,可以具有更强的结合亲和力、增强的降低、抑制或中和动物中ENO1活性的能力,和/或增强的在基于细胞的测定中抑制ENO1介导的信号传导的能力。
本发明提供了如下任一项所述的物质:
(1)核酸分子,所述核酸分子编码上述单克隆抗体或其功能片段;
(2)载体,所述载体包括(1)中所述的核酸分子;
(3)宿主细胞,所述宿主细胞包括(1)中所述的核酸分子或(2)中所述的载体;
(4)药物偶联物,所述药物偶联物包括上述单克隆抗体或其功能片段;
(5)一种检测ENO1的产品,所述产品包括上述单克隆抗体或其功能片段。
在本发明中,载体是指可以将编码蛋白质的多核苷酸可操作地插入其中以引起所述蛋白质表达的媒介物。载体可用于转化、转导或转染宿主细胞,以使其在宿主细胞内表达携带的遗传元件。载体可以含有多种用于控制表达的元件,包含启动子序列、转录起始序列、增强子序列、选择元件和报告基因。另外,载体可以含有复制起点。术语复制起点是指当在载体中存在时其起始复制的序列。复制起点可被复制起始因子或替代地被DNA解螺旋酶识别。载体还可以包括有助于其进入细胞的材料,包括但不限于病毒颗粒、脂质体或蛋白质包膜。
载体可以是重组表达载体或克隆载体。本发明提供了载体(例如表达载体),其含有编码抗ENO1中和抗体的本发明提供的核酸序列、至少一个与核酸序列可操作地连接的启动子和/或至少一个选择标记。载体的实例包括但不限于逆转录病毒(包括慢病毒)、腺病毒、腺相关病毒、疱疹病毒(例如单纯疱疹病毒)、痘病毒、杆状病毒、乳头状瘤病毒、乳多空病毒(例如SV40)、λ噬菌体和M13噬菌体、质粒,包括但不限于pcDNA3.3、pMD18-T、pOptivec、pCMV、pEGFP、pIRES、pQD-Hyg-GSeu、pALTER、pBAD、pcDNA、pCal、pL、pET、pGEMEX、pGEX、pCI、pEGFT、pSV2、pFUSE、pVITRO、pVIVO、pMAL、pMONO、pSELECT、pUNO、pDUO、Psg5L、pBABE、pWPXL、pBI、p15TV-L、pPro18、pTD、pRS10、pLexA、pACT2.2、pCMV-SCRIPT.RTM.、pCDM8、pCDNA1.1/amp、pcDNA3.1、pRc/RSV、PCR2.1、pEF-1、pFB、pSG5、pXT1、pCDEF3、pSVSPORT、pEF-Bos。
在本发明中,宿主细胞实际上可以是表达载体可用的任何细胞。包括原核细胞、真核细胞,所述原核细胞包括但不限于真细菌,如革兰氏阴性或革兰氏阳性生物体,例如肠内菌科(Enterobacteriaceae),如埃希氏菌属(Escherichia),例如大肠杆菌;肠杆菌属(Enterobacter);欧文氏菌属(Erwinia);克雷伯氏菌属(Klebsiella);变形杆菌属(Proteus);沙门氏菌属(Salmonella),例如鼠伤寒沙门氏菌(Salmonella typhimurium);沙雷氏菌属(Serratia),例如粘质沙雷氏菌(Serratia marcescans);和志贺杆菌属(Shigella),以及芽孢杆菌属(Bacilli),如枯草芽孢杆菌(B.subtilis)和地衣芽孢杆菌(B.licheniformis);假单胞菌属(Pseudomonas),如绿脓杆菌(P.aeruginosa);和链霉菌属(Streptomyces)。
真核细胞包括但不限于原生生物细胞、动物细胞、植物细胞或真菌细胞,所述动物细胞包括哺乳动物细胞、鸟类细胞、昆虫细胞;其中,哺乳动物细胞包括但不限于中国仓鼠卵巢细胞、SP2/0细胞、HeLa细胞、幼仓鼠肾细胞、癌细胞。
本发明提供了一种衍生物,所述衍生物包括在上述单克隆抗体或上述核酸分子上连接诊断试剂、治疗试剂;
所述诊断试剂在本领域中是已知的,其与蛋白质(包括抗体)连接的方法同样是已知的,包括但不限于荧光标记、放射性同位素、化学发光分子、顺磁性离子或自旋捕获试剂。
其中,荧光标记包括但不限于Alexa 350、Alexa 430、AMCA、BODIPY 630/650、BODIPY 650/665、BODIPY-FL、BODIPY-R6G、BODIPY-TMR、BODIPY-TRX、Cascade Blue、Cy3、Cy5,6-FAM、异硫氰酸荧光素、HEX、6-JOE、Oregon Green488、Oregon Green 500、OregonGreen 514、Pacific Blue、REG、罗丹明绿、罗丹明红、Renographin、ROX、TAMRA、TET、四甲基罗丹明和/或Texas Red。
放射性同位素包括但不限于砹211、14碳、51铬、36氯、57钴、58钴、铜67、152Eu、镓67、3氢、碘123、碘125、碘131、铟111、59铁、32磷、铼186、铼188、75硒、35硫、锝99m(technicium)和/或钇90。
顺磁性离子包括但不限于铬(III)、锰(II)、铁(III)、铁(II)、钴(II)、镍(II)、铜(II)、钕(III)、钐(III)、镱(III)、钆(III)、钒(II)、铽(III)、镝(III)、钬(III)和/或铒(III)的离子。
所述治疗试剂包括但不限于细胞因子、化学治疗剂、放射治疗剂、抗炎剂、激素、抗体Fc片段、TLR激动剂、含CpG的分子或免疫共刺激分子。
其中,细胞因子包括但不限于IL-1、IL-2、IL-3、IL-4、IL-5、IL-6、IL-7、IL-8、IL-9、IL-10、IL-11、IL-12、IL-13、IL-14、IL-15、IL-16、IL-17、IL-18、IL-19、IL-20、IL-21、IL-22、IL-23、TNFα、GM-CSF、INFα、IFNβ和IFNγ。
化学治疗剂包括但不限于顺铂(CDDP)、卡铂、甲基苄肼、氮芥、环磷酰胺、喜树碱、异环磷酰胺、美法仑(melphalan)、苯丁酸氮芥、白消安(busulfan)、亚硝基脲(nitrosurea)、更生霉素、柔红霉素(daunorubicin)、阿霉素(doxorubicin)、博来霉素(bleomycin)、普卡霉素(plicomycin)、丝裂霉素、依托泊甙(VP 16)、他莫昔芬、雷洛昔芬、雌激素受体结合剂、紫杉醇、吉西他滨(gemcitabien)、诺维本(navelbine)、法尼基蛋白(farnesyl-protein)转移酶抑制剂、反铂(transplatinum)、5-氟脲嘧啶、长春新碱、长春花碱和甲氨蝶呤。
另一类治疗剂是毒素,包括但不限于霍乱毒素、肉毒杆菌毒素、百日咳毒素、蓖麻毒蛋白A和B链以及其他天然毒素和合成毒素。
抗炎剂包括但不限于NSAID、类固醇类、瑞帕霉素、英夫利昔单抗和地尼白介素(ontak)。
TLR激动剂是刺激或打开免疫系统的化合物。TLR9的天然激动剂是细菌和病毒中常见的DNA的组分。TLR7和TLR8的天然激动剂是病毒中发现的RNA模式。在识别其天然DNA和RNA激动剂后,TLR7、8和9各自引发不同级联的保护性免疫应答。TLR激动剂包括寡脱氧核苷酸、透明质酸片段、咪喹莫特、薰草菌素C、脂质A、loroxibine、LPS、单磷酰脂质A(monophosphoryl lipda A)、肉豆蔻醚、雷西莫特(resiquimod)、鼠伤寒沙门氏菌(S.typhimurium)鞭毛蛋白、HKLM、PAM3CSK4和聚肌胞苷酸(polyI:C)。
本发明提供了一种药物组合物,所述药物组合物包括上述单克隆抗体、上述物质或上述衍生物。
所述药物组合物还包括免疫调节剂。
在本发明中,免疫调节剂是指调节免疫应答的物质、试剂、信号传导途径或其组分。免疫应答的调节、修饰或调制是指免疫细胞自身或其细胞活性发生改变。这种调节包括对免疫系统的刺激或抑制,其表现为各种细胞类型数量的增加或减少,以及这些免疫细胞活力的增强或降低,或免疫系统内可能发生的其他变化。
所述药物组合物还包括药学上可接受的载体。
在本发明中,药学上可接受的是指该组合物足以实现治疗效果并无不良副作用,此外,可以根据疾病的类型,患者的年龄,体重,健康状况,性别和药物敏感性较容易地确定给药途径,给药方式,给药频率,疗程,与本发明公开的组合物结合或联合使用的药物,以及医学中已知的其他因素。药学上可接受的载体包括但不限于粘合剂,润滑剂,崩解剂,赋形剂,增溶剂,分散剂,稳定剂,悬浮剂,着色剂和食用香料。对于注射制剂可包括缓冲剂,防腐剂,镇痛剂,增溶剂,等渗剂和稳定剂。对于局部给药的制剂,载体可包括基础物,赋形剂,润滑剂和防腐剂。
本发明所公开的组合物可以与上述药学上可接受的载体组合配制成各种剂型。例如,对于口服给药,可以将药物组合物配制成片剂,锭剂,胶囊,酏剂,悬浮液,糖浆或糯米纸囊剂。对于可注射制剂,可将药物组合物置于可作为单次剂量或多次剂量容器的安瓿瓶中。药物组合物还可以配制成溶液,悬浮液,片剂,丸剂,胶囊和长效制剂。
另一方面,适用于药物制剂的载体,赋形剂和稀释剂的实例包括但不限于乳糖,右旋糖,蔗糖,山梨糖醇,甘露醇,木糖醇,赤藓糖醇,麦芽糖醇,淀粉,阿拉伯胶,海藻酸盐,明胶,磷酸钙,硅酸钙,纤维素,甲基纤维素,微晶纤维素,聚乙烯吡咯烷酮,水,羟基苯甲酸甲酯,羟基苯甲酸丙酯,滑石,硬脂酸镁和矿物油。此外,药学制剂可以进一步地包括填充剂,抗凝剂,润滑剂,保湿剂,调味剂和防腐剂。
本发明提供了上述单克隆抗体、上述物质、上述衍生物或上述药物组合物在制备预防和/或治疗ENO1相关疾病的药物中的应用。
ENO1相关疾病是存在表达ENO1的细胞的疾病或病症。
所述ENO1相关疾病包括肿瘤、代谢性疾病和/或免疫疾病。
其中,肿瘤包括但不限于肝癌、胃癌、肾癌、乳腺癌、结直肠癌、头颈癌、子宫内膜癌。
免疫疾病包括但不限于类风湿性关节炎、骨关节炎、系统性红斑性狼疮、哮喘、炎症性肠道疾病、多发性硬化、克罗恩氏病、胃炎、桥本氏甲状腺炎、强直性脊柱炎和移植物对抗宿主疾病。
代谢疾病包括但不限于骨质疏松、糖尿病、高血脂症、痛风、维生素D缺乏病、蛋白质-能量营养不良症。
下面结合具体实施例进一步阐述此发明。应理解的是,在此描述的特定实施方式通过举例的方式来表示,并不作为对本发明的限制。在不偏离本发明范围的情况下,本发明的主要特征可以用于各种实施方式。
实施例1ENO1蛋白的表达和纯化
1、ENO1蛋白的表达
用接种环直接取冻存的大肠杆菌EN0-1/Rosetta,在LB培养基平板表面划线,于37℃培养16小时,摇床37℃、300rpm。挑取一个单菌落,转到3ml LB培养基中,37℃,280rpm,培养3-5小时,测得OD600=0.6-0.7,SDS-PAGE实验证明筛选最佳的IPTG诱导浓度比例1:6000和诱导时间3h(图1)。
2、ENO1蛋白的纯化
Ni金属螯合亲和柱的挂Ni、预平衡处理,变性蛋白加入适宜体积Ni-NTA His-BindResins(3-5mg/ml柱容量,保守地可按2mg/ml估算,按蛋白含量估算),室温颠倒地结合1h,装柱,收集流穿液备检。取部分蛋白,确定洗涤条件和洗脱的pH值。采用0、20、50、500mM洗脱一次,直到无蛋白洗脱。所有洗脱液洗脱后立即调节pH至8.0,收集所有流穿液SDS-PAGE分析,实验结果表明:0、500mM咪唑洗脱,循环上样3h效果好(图2)。
实施例2ENO1mAb的制备
1、动物免疫
将ENO1蛋白(100μg/ml)与弗氏完全佐剂等体积混合,然后经背部皮下多点注射雌性BALB/c小鼠;间隔三周后进行二次免疫,将ENO1蛋白(200μg/ml)与弗氏不完全佐剂等体积混匀,在BALB/c小鼠背部皮下多点注射;三周后进行三次免疫,与第二次免疫方案相同,三周后进行四次免疫,与第三次免疫方案相同,采集BALB/c小鼠的血液,应用间接ELISA法,测定血清中抗体滴度。冲击免疫为中(500μg/ml)ENO1静脉注射。
2、BALB-C小鼠胸腺细胞(滋养细胞)
将小鼠从75%酒精中取出,用眼科剪剪开小鼠腹部的皮肤,暴露腹部取脾脏,取出完整脾脏,剪去其周围的脂肪或组织,拍照。将胸腺在100目筛网上用无齿镊轻轻研磨,用培养基冲洗细胞过筛网到大平皿中,获得胸腺细胞悬液,将细胞悬液转移至50ml离心管中。离心、洗涤:1200rpm,5min,用移液管吸弃上清,加入20ml ImEm培养液轻轻吹匀,重悬。
3、免疫后小鼠脾细胞
用眼科剪剪开小鼠腹部的皮肤,暴露腹部取脾脏,用另一眼科剪剪开小鼠腹膜,取出完整脾脏,剪去其周围的脂肪或组织。将脾脏在100目筛网上用无齿镊轻轻摩擦,用培养基冲洗细胞过筛网到大平皿中,获得脾脏细胞悬液,将细胞悬液转移至50ml离心管中,取20μl细胞悬液进行细胞计数。
4、SP2/0细胞培养
用移液管弃去DmEm-H培养液,加入10ml ImDm培养液浸泡,轻轻拍打培养瓶使细胞脱落,收集细胞悬液至50ml离心管,取20μl细胞计数,离心、洗涤:1200rpm,5min,用移液管吸弃上清,加入20ml培养液轻轻吹匀,重悬。
5、脾细胞与SP2/0细胞混合
SP2/0的细胞悬液转移至已弃上清的脾细胞离心管中轻轻吹匀,重悬脾细胞与SP2/0混合:1200rpm,5min,离心,用移液管吸弃上清,将50ml离心管于掌中轻轻摩擦至底部细胞充分混合均匀,准备45℃温水于保温瓶中,用前加入些许冰块,调整温度至38℃,将50ml离心管至于温水中,缓缓加入1ml PEG,边加边轻轻摇匀,随后缓缓加入10ml ImDm培养液,合计5min。离心:1200rpm,5min,移液管吸弃上清,加入胸腺细胞悬液,轻轻混合均匀,1200rpm,5min,离心,用移液管吸弃上清,加入24ml ImDm培养液,重悬。将混合细胞与培养基(2ml细胞悬液+1ml HAT+2ml ImDm培养液+10ml HyClone高级进口血清+25ml 2.5%甲基纤维素),上下颠倒50ml离心管20min,至管内完全均匀。
6、阳性克隆的筛选与克隆化
待克隆化的阳性融合细胞,可先转入24孔板扩大培养后再克隆,也可边扩大边从96孔板取样直接克隆。用吸管将细胞群吹打分散,制成悬液,按前述方法精确计数活细胞数。从96孔板直接取样克隆,计数取样只用1滴悬液,用另一支滴管加无血清培养液9滴作10倍稀释,计数。用HT培养液将细胞稀释至每毫升5、30和50个细胞各40ml。每种杂交瘤细胞用一块备好的含饲养细胞的96孔板,每个稀释度32孔,每孔加细胞悬液0.1ml,每孔应含0.5、3和5个细胞。由于计数和加样的误差,往往偏高或偏低。如从96孔板直接取细胞克隆时,应在克隆完后立即把孔中和稀释管中剩余细胞全部转入24孔中扩大培养,以备失败时重新克隆。在37℃,5%-7%培养箱中培养7天左右,镜检,标出所有板孔的细胞群落数。采样,进行抗体检测。将单克隆阳性孔细胞先转入24孔,后在培养瓶中扩大培养,然后冻存。共获得4株杂交瘤细胞株,分别命名为ENO1mAb(17G12、12A11、21A3和23G5),随后,进行杂交瘤细胞的冻存。
7、杂交瘤细胞的培养
杂交瘤细胞以含有12%FBS、2mmol/L L-谷氨酰胺、100U/mL青霉素及100μg/mL链霉素的DMEM-H完全培养液,在37℃、5%CO2,大批量培养杂交瘤细胞收集上清时,一般稳定传代15-20次后,需重新复苏一批新细胞;一般使用1:2比例传代,于传代后24h换液,48h后长满再传代。
8、无血清杂交瘤上清的收集
杂交瘤扩大培养至T175瓶中,每次约30瓶;传代后第2天换液,48h后长到约75-90%满后,移去完全培养基,改换无血清培养基约20ml/瓶;次日每天用1mol/L的无菌NaOH调节液体的PH至7.0-7.5左右,并添加新鲜无血清培养基2-3ml;待贴壁细胞逐渐减少,细胞活度下降至40%-50%时,3000rpm,10min离心,收集无血清杂交瘤上清。
9、抗体纯化
将预装柱上面盖子旋开,用蠕动泵滴PBS赶走气泡,PBS平衡大约20个柱体积。上完样后取1ml的流穿于1.5ml的EP管中,用PBS洗杂,调流速至1ml/min(1ml柱子),洗杂至少5个柱体积,即5ml PBS,测OD280;管中加入75μl的1MTris-HCl(pH 9.0)中和buffer。用0.1MGlycine-HCl(pH 2.7)洗脱buffer洗脱柱子,调流速1ml/min,接洗脱下来的液体,立即混合,使洗脱下来的抗体立即恢复PH值至7.0左右,每管测OD280。当OD值小于0.1时则可以停止接液。将洗脱下来的抗体混合,将混合后蛋白取10μl留样;将浓缩好的抗体放入20倍体积的预先预冷的PBS中4℃,搅拌透析,每次透析4h,透析3次;透析好的抗体离心,12000rpm,离心15min。0.22μm滤器过滤除菌,分装,以避免反复冻融。将收集到的ENO1mAb进行抗体浓度测定及SDS-PAGE电泳分析。经SDSPAGE验证显示,ENO1mAb纯化效果较好,无明显杂带,符合抗体重链和轻链大小(图3)。
10、抗体亚类的检测
用PBS将捕获抗体稀释为1μg/ml,100μl/孔加入96孔酶标板,4℃孵育16h;弃去包被抗体,加入含1%BSA的PBS封闭液,300μl/孔,37℃封闭酶标板4h;弃去封闭液,用含0.05%Tween-20的PBS洗板三次,300μl/孔,1min/次×5次;按设计加入待测样品,100μl/孔,37℃孵育1.5h;分别加入HRP标记的各种类型二抗(HRP标记的羊抗鼠IgM、IgG1、IgG2a、IgG2b、IgG3、IgA、κorλ,1:2000稀释),100μl/孔,37℃孵育1h;PBS再洗涤2次;加入TMB显色,100μl/孔,37℃孵育30min;加入2M H2SO4,50μl/孔。酶标仪测定四种抗体(17G12、12A11、21A3、23G5)亚型,其对应的亚型为(IgG2b、IgG1、IgG1、IgG2a)(图4)。
11、非竞争性ELISA检测抗ENO1单抗对重组ENO1的亲和力
在采用非竞争性ELISA法(Beatty法)可测定各单抗的亲和常数Ka。首先优化ENO1抗原包被浓度梯度和抗体系列稀释的梯度浓度,最终确定正式试验条件为:以不同浓度梯度的ENO1抗原(0.593、0.395、0.2633、0.1755、0.117、0.078、0.000g/ml)包被,采用四种单抗稀释浓度(32000、16000、8000、4000、2000、1000、500、250、125、62.5、31.2、15.6、7.8、3.9、1.95、0.98、0.5、0.25、0.12、0.06、0.03、0.0125、0ng/ml)。此条件下针对未标记的ENO1单抗进行测定,绘制抗体OD值为纵坐标,抗体浓度对数为横坐标的反应曲线,可以在各个ENO1包被浓度下均能得到较好的典型S型反应曲线,计算出待检测抗体解离常数KD,其中,23G5KD=3.729x10-9(图5)。
12、ENO1单抗23G5测序
采用培养23G5杂交瘤细胞总RNA提取试剂盒进行RNA的抽提,提取的RNA溶液于-70℃保存。对提取的RNA进行反转录,末端修复,然后进行接头连接和PCR扩增纯化得到NGS文库,文库库检合格后用于上机测序。对构建好的文库进行高通量测序,每个样品测6G数据。二代高通量测序仪器测序得到的原始图像数据经base calling转化为序列数据,称之为raw data或raw reads,结果以FASTQ文件格式存储,包含reads的序列以及碱基的测序质量。对raw data进行数据过滤,从而获得clean data用于后续分析。使用Trinity-v2.11.0软件对Clean Reads进行无参组装。使用igblast进行比对分析,小鼠的免疫数据库来自IMGT。然后对fmt7文件进行整理和过滤。使用Change-O的子模MakeDb.py,ParseDb.py及ConvertDb.py进行分析。对数据进行分析整理得到单克隆抗体轻链和重链的序列,并根据序列得到可变区氨基酸序列,23G5抗体的序列如表1所示。
表1抗体序列
/>
/>
实施例3ENO1单抗23G5对骨关节炎小鼠的治疗效果研究
膝盖前交叉韧带切除术(Anterior cruciate ligament transection,ACLT)的骨关节炎小鼠模型的构建:小鼠用右美托咪啶(0.25mg/kg)和氯胺酮(25mg/kg)肌内注射麻醉,常规备皮消毒左侧膝关节,取髌旁内侧切口显露膝关节,显露髌骨后将髌骨向外侧脱位,尽量屈曲膝关节显露ACLT,在直视下切断前交叉韧带,彻底止血,将髌骨复位,1号丝线逐层闭合关节腔。假手术组仅切开缝合左侧膝关节,未切断前十字韧带。
6周龄雄性C57B6N小鼠(n=25)购置于北京维通利华实验动物技术有限公司。经2周适应环境后,将小鼠随机分为5组,每组5只:①正常组,未腹腔注射;②假手术组,未腹腔注射;③低剂量23G5单抗ACLT模型组,ACLT后腹腔注射单抗23G5(2.5mg/kg);④中剂量23G5单抗ACLT模型组,ACLT后腹腔注射单抗23G5(10mg/kg);⑤高剂量23G5单抗ACLT模型组,ACLT后腹腔注射单抗23G5(40mg/kg)。
治疗7周后,micro-CT扫描显示:与对照组相比,其通过腹膜内注射不同剂量(2.5、10和40mg/kg)后小鼠左侧胫骨平台的软骨下骨显著改善(图6)。此外,Micro-CT定量分析显示,高、中和低剂量23G5单抗对骨密度(BMD)、骨体积/组织体积分数(BV/TV)、孔隙率(PO)、结构模型指数(SMI)、骨小梁数目(Tb.N)、骨小梁厚度(Tb.Th)、骨小梁分离度(Tb.Sp)和骨小梁模式因素(Tb.Pf)发挥拮抗作用(图7)。进一步通过HE和番红O染色以评价单抗23G5对小鼠胫骨内侧平台关节软骨的影响。与对照组相比,腹腔注射单抗23G5(2.5、10和40mg/kg)显著减少关节软骨的破坏(图8)。
实施例4ENO1单抗23G5对骨质疏松大鼠的治疗效果研究
骨质疏松大鼠模型(OVX)的建立:40只10周龄SD雌性大鼠(体重230-250g),给予充足的食物和水,适应性喂养一周后,所有大鼠接受异氟烷吸入,联合腹腔注射10%水合氯醛(300mg/kg)麻醉。无菌条件下,30只大鼠接受双侧卵巢切除手术(OVX),构建去卵巢诱导的骨质疏松动物模型;10只大鼠切除双侧卵巢周围同体积脂肪组织(假手术,Sham)。
将40只大鼠分为5组:①假手术组(n=10):Sham大鼠接受100μl PBS治疗;②OVX组(n=10):OVX大鼠接受100μl PBS治疗;③OVX+低剂量组(n=10):OVX大鼠接受低剂量23G5单抗治疗(2.5mg/kg比例的23G5单抗溶于100μl PBS);④OVX+中剂量组(n=10):OVX大鼠接受中剂量23G5单抗治疗(10mg/kg比例的23G5单抗溶于100μl PBS;⑤OVX+高剂量组(n=10):OVX大鼠接受高剂量23G5单抗治疗(40mg/kg比例的23G5单抗溶于100μl PBS)。所有大鼠通过腹腔注射,每周1次,持续8周。
治疗8周后,micro-CT扫描显示:OVX组(PBS处理)股骨远端的骨小梁形态中断、稀疏;而OVX+23G5组中随着23G5抗体浓度逐渐增加后,这种糟糕的状况获得了明显改善(图9)。
此外,相较于PBS处理组的OVX大鼠,OVX+23G5组处理组的大鼠相关参数包括骨体积/组织体积分数(BV/TV)、骨小梁数目(Tb.N)、骨小梁厚度(Tb.Th)和骨密度(BMD)的值明显提高;相反,骨小梁分离度(Tb.Sp)的值明显降低(图10)。
实施例5ENO1单抗23G5对类风湿关节炎大鼠的治疗效果研究
选取8-12周龄Wistar大鼠,体重180±20g,购自北京维通利华实验动物技术有限公司。
类风湿关节炎(CIA)大鼠构建方法如下:乳化:将鸡II型胶原(4mg/ml)和完全弗式佐剂(CFA)(4mg/ml)1:1进行混合,用10ml针筒反复抽吸,充分乳化(至少30分钟),数小时后,滴少许乳化剂到水面上,如果30s乳剂不会迅速扩展,则为乳化完全。首次免疫:将乳化剂转移至1ml注射器,在距离Wistar大鼠尾根部2cm处,避开血管,进行多点皮下注射,每只大鼠共注射乳化剂100μl。记作第0天。二次免疫:首次免疫7d后,进行加强免疫,将鸡II型胶原(4mg/ml)和非完全弗式佐剂(4mg/ml)1:1进行混合,具体方法同上。
在第33d,36d,39d,42d,48d和51d时,23G5单抗处理组CIA大鼠的关节炎评分低于Model组(p<0.05),说明CIA大鼠体内被23G5单抗作用后,CIA大鼠关节炎症状减轻,同时,随着23G5单抗作用的剂量的提高,CIA大鼠关节炎症状减轻,说明ENO1单抗能降低CIA大鼠炎症的抑制作用并且显示剂量依赖(图11)。
Model组中CIA大鼠滑膜组织中炎症因子TNF-α、IL-1β、IL-6和IL-17分泌增高,23G5单抗处理后CIA大鼠滑膜组织中炎症因子TNF-α、IL-1β、IL-6和IL-17mRNA表达量显著低于Model组(p<0.01或0.05),提示CIA大鼠体内被23G5单抗作用后,CIA大鼠关节滑膜组织炎症因子TNF-α、IL-1β、IL-6和IL-17分泌减少。提示23G5单抗能抑制CIA大鼠滑膜组织分泌TNF-α、IL-1β、IL-6和IL-17分泌减少,且有剂量依赖作用(图12)。
上述实施例的说明只是用于理解本发明的方法及其核心思想。应当指出,对于本领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以对本发明进行若干改进和修饰,这些改进和修饰也将落入本发明权利要求的保护范围内。
Claims (10)
1.一种抗ENO1的单克隆抗体,其特征在于,所述单克隆抗体包括具有与SEQ ID NO:1、2、3所示的氨基酸序列至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的重链可变区HCDR1、HCDR2、HCDR3,或其具有一个或多个保守氨基酸取代的任何变体;
以及具有与SEQ ID NO:9、10、11所示的氨基酸序列至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的轻链可变区LCDR1、LCDR2、LCDR3,或其具有一个或多个保守氨基酸取代的任何变体;
优选的,所述HCDR1、HCDR2、HCDR3的氨基酸序列分别如SEQ ID NO:1、2、3所示;
所述LCDR1、LCDR2、LCDR3的氨基酸序列分别如SEQ ID NO:9、10、11所示。
2.根据权利要求1所述的单克隆抗体,其特征在于,所述单克隆抗体还包括具有与SEQID NO:4、5、6、7所示的氨基酸序列至少90%、至少92%、至少93%、至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的重链可变区框架区HFR1、HFR2、HFR3、HFR4,或其具有一个或多个保守氨基酸取代的任何变体;
以及具有与SEQ ID NO:12、13、14、15所示的氨基酸序列至少90%、至少92%、至少93%、至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的轻链可变区框架区LFR1、LFR2、LFR3、LFR4,或其具有一个或多个保守氨基酸取代的任何变体;
优选的,所述重链可变区框架区HFR1、HFR2、HFR3、HFR4的氨基酸序列分别如SEQ IDNO:4、5、6、7所示;
所述轻链可变区框架区LFR1、LFR2、LFR3、LFR4的氨基酸序列分别如SEQ ID NO:12、13、14、15所示;
优选的,所述单克隆抗体重链可变区具有与SEQ ID NO:8所示的氨基酸序列至少70%、至少75%、至少80%、至少85%、至少90%、至少92%、至少93%、95%、至少96%、至少97%、至少98%、至少99%序列同一性的VH,或其具有一个或多个保守氨基酸取代的任何变体;
所述轻链可变区具有与SEQ ID NO:16所示的氨基酸序列至少70%、至少75%、至少80%、至少85%、至少90%、至少92%、至少93%、95%、至少96%、至少97%、至少98%、至少99%序列同一性的VL,或其具有一个或多个保守氨基酸取代的任何变体;
优选的,所述单克隆抗体的VH具有如SEQ ID NO:8所示的氨基酸序列,所述单克隆抗体的VL具有如SEQ ID NO:16所示的氨基酸序列;
优选的,所述单克隆抗体还包括抗体重链恒定区和/或抗体轻链恒定区的全部或者部分;
优选的,所述单克隆抗体的重链亚型为IgG2;
优选的,所述IgG2包括IgG2a、IgG2b;
优选的,所述单克隆抗体的重链亚型为IgG2a;
优选的,所述单克隆抗体的轻链亚型为κ。
3.如下任一项所述的物质:
(1)核酸分子,其特征在于,所述核酸分子编码权利要求1或2所述的单克隆抗体或其功能片段;
(2)载体,其特征在于,所述载体包括(1)中所述的核酸分子;
(3)宿主细胞,其特征在于,所述宿主细胞包括(1)中所述的核酸分子或(2)中所述的载体;
(4)药物偶联物,其特征在于,所述药物偶联物包括权利要求1或2所述的单克隆抗体或其功能片段;
(5)一种检测ENO1的产品,其特征在于,所述产品包括权利要求1或2所述的单克隆抗体或其功能片段。
4.根据权利要求3所述的物质,其特征在于,(1)中编码HCDR1、HCDR2、HCDR3的核酸分子具有与SEQ ID NO:17、18、19所示的核苷酸序列至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列;
编码LCDR1、LCDR2、LCDR3的核酸分子具有与SEQ ID NO:25、26、27所示的核苷酸序列至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列;
优选的,编码HCDR1、HCDR2、HCDR3的核酸分子的核苷酸序列分别如SEQ ID NO:17、18、19所示,编码LCDR1、LCDR2、LCDR3的核酸分子的核苷酸序列分别如SEQ ID NO:25、26、27所示;
优选的,编码HFR1、HFR2、HFR3、HFR4的核酸分子具有与SEQ ID NO:20、21、22、23所示的核苷酸序列至少90%、至少92%、至少93%、至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列;
编码LFR1、LFR2、LFR3、LFR4的核酸分子具有与SEQ ID NO:28、29、30、31所示的核苷酸序列至少90%、至少92%、至少93%、至少95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列;
优选的,编码HFR1、HFR2、HFR3、HFR4的核酸分子的核苷酸序列分别如SEQ ID NO:20、21、22、23所示,编码LFR1、LFR2、LFR3、LFR4的核酸分子的核苷酸序列分别如SEQ ID NO:28、29、30、31所示;
优选的,编码VH的核酸分子具有与SEQ ID NO:24所示的核苷酸序列至少70%、至少75%、至少80%、至少85%、至少90%、至少92%、至少93%、95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列;
编码VL的核酸分子具有与SEQ ID NO:32所示的核苷酸序列至少70%、至少75%、至少80%、至少85%、至少90%、至少92%、至少93%、95%、至少96%、至少97%、至少98%、至少99%序列同一性的核苷酸序列;
优选的,编码VH的核酸分子的核苷酸序列如SEQ ID NO:24所示,编码VL的核酸分子的核苷酸序列如SEQ ID NO:32所示。
5.根据权利要求3所述的物质,其特征在于,(2)中所述的载体具有与抗体可操作的连接的信号肽;
优选的,所述载体还包括用于控制表达的元件;
优选的,所述载体还包括有助于其进入细胞的材料;
优选的,(3)中所述的宿主细胞包括原核细胞、真核细胞;
优选的,所述原核细胞包括大肠杆菌;
优选的,所述真核细胞包括原生生物细胞、动物细胞、植物细胞或真菌细胞;
优选的,所述动物细胞包括哺乳动物细胞、鸟类细胞、昆虫细胞。
6.根据权利要求3所述的物质,其特征在于,(4)中所述的药物偶联物还包括可检测标记物、药物、毒素、细胞因子和/或酶;
优选的,(5)中所述的产品还包括用于执行抗原-抗体反应的试剂或用于检测反应的试剂;
优选的,用于执行抗原-抗体反应的试剂包括缓冲剂、盐;
优选的,所述产品包括试剂盒、试纸条;
优选的,所述试剂盒包括连接有信号产生化合物的抗体的缀合物。
7.一种衍生物,其特征在于,所述衍生物包括在权利要求1或2所述的单克隆抗体或权利要求3或4所述的核酸分子上连接诊断试剂、治疗试剂;
优选的,所述诊断试剂包括荧光标记、放射性同位素、化学发光分子、顺磁性离子或自旋捕获试剂;
优选的,所述治疗试剂包括细胞因子、化学治疗剂、放射治疗剂、抗炎剂、激素、抗体Fc片段、TLR激动剂、含CpG的分子或免疫共刺激分子。
8.如下任一项所述的方法:
(1)一种制备权利要求1或2所述的单克隆抗体的方法,其特征在于,所述方法包括:培养权利要求3-6任一项所述的宿主细胞,回收单克隆抗体;
(2)一种检测样本中ENO1的方法,其特征在于,所述方法包括:使权利要求1或2所述的抗ENO1的单克隆抗体与待测样本接触,从而检测待测样本中ENO1的水平;
优选的,(1)中所述的方法还包括纯化所述的单克隆抗体。
9.一种药物组合物,其特征在于,所述药物组合物包括权利要求1或2所述的单克隆抗体、权利要求3-6任一项所述的物质或权利要求7所述的衍生物;
优选的,所述药物组合物还包括免疫调节剂;
优选的,所述药物组合物还包括药学上可接受的载体。
10.如下任一项所述的应用:
(1)权利要求1或2所述的单克隆抗体、权利要求3-6任一项所述的物质或权利要求7所述的衍生物在检测ENO1中的应用;
(2)权利要求1或2所述的单克隆抗体、权利要求3-6任一项所述的物质或权利要求7所述的衍生物在制备诊断ENO1相关疾病的产品中的应用;
(3)权利要求1或2所述的单克隆抗体、权利要求3-6任一项所述的物质、权利要求7所述的衍生物或权利要求9所述的药物组合物在制备预防和/或治疗ENO1相关疾病的药物中的应用;
优选的,所述ENO1相关疾病包括肿瘤、代谢性疾病和/或免疫疾病;
优选的,所述ENO1相关疾病选自代谢性疾病和/或免疫疾病;
优选的,所述免疫疾病包括骨关节炎、类风湿关节炎;
优选的,所述代谢性疾病选自骨质疏松。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310510154.4A CN117264066B (zh) | 2023-05-08 | 2023-05-08 | 一种eno1抗体及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310510154.4A CN117264066B (zh) | 2023-05-08 | 2023-05-08 | 一种eno1抗体及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN117264066A true CN117264066A (zh) | 2023-12-22 |
CN117264066B CN117264066B (zh) | 2024-03-22 |
Family
ID=89206969
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310510154.4A Active CN117264066B (zh) | 2023-05-08 | 2023-05-08 | 一种eno1抗体及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN117264066B (zh) |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20190322762A1 (en) * | 2018-03-16 | 2019-10-24 | Development Center For Biotechnology | Antibodies specific to alpha-enolase and uses thereof |
CN112111462A (zh) * | 2020-09-14 | 2020-12-22 | 兰州大学 | 烯醇化酶eno1单克隆抗体及其应用 |
US20210221847A1 (en) * | 2018-07-03 | 2021-07-22 | Seoul National University R&Db Foundation | Peptide for treating rheumatoid arthritis and use thereof |
CN114426581A (zh) * | 2022-01-07 | 2022-05-03 | 陕西脉元生物科技有限公司 | 一种烯醇化酶单克隆抗体及其制备方法和应用 |
CN114478784A (zh) * | 2022-01-07 | 2022-05-13 | 陕西脉元生物科技有限公司 | 一种eno2单克隆抗体1c5及其制备方法和应用 |
CN116333150A (zh) * | 2023-05-08 | 2023-06-27 | 北京积水潭医院 | 一种eno1抗体及其在治疗肿瘤中的应用 |
-
2023
- 2023-05-08 CN CN202310510154.4A patent/CN117264066B/zh active Active
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20190322762A1 (en) * | 2018-03-16 | 2019-10-24 | Development Center For Biotechnology | Antibodies specific to alpha-enolase and uses thereof |
US20210221847A1 (en) * | 2018-07-03 | 2021-07-22 | Seoul National University R&Db Foundation | Peptide for treating rheumatoid arthritis and use thereof |
CN112111462A (zh) * | 2020-09-14 | 2020-12-22 | 兰州大学 | 烯醇化酶eno1单克隆抗体及其应用 |
CN114426581A (zh) * | 2022-01-07 | 2022-05-03 | 陕西脉元生物科技有限公司 | 一种烯醇化酶单克隆抗体及其制备方法和应用 |
CN114478784A (zh) * | 2022-01-07 | 2022-05-13 | 陕西脉元生物科技有限公司 | 一种eno2单克隆抗体1c5及其制备方法和应用 |
CN116333150A (zh) * | 2023-05-08 | 2023-06-27 | 北京积水潭医院 | 一种eno1抗体及其在治疗肿瘤中的应用 |
Non-Patent Citations (2)
Title |
---|
王珍妮等: "α-烯醇化酶的翻译后修饰对类风湿关节炎免疫诊断价值", 现代免疫学, vol. 38, no. 5, 30 September 2018 (2018-09-30), pages 353 - 359 * |
肖翠等: "糖酵解酶在类风湿关节炎发病机制中的研究进展", 实用临床医学, vol. 21, no. 4, 20 April 2020 (2020-04-20), pages 100 - 103 * |
Also Published As
Publication number | Publication date |
---|---|
CN117264066B (zh) | 2024-03-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20140205611A1 (en) | Monoclonal antibodies that bind to hgm-csf and medical compositions comprising same | |
KR102599907B1 (ko) | 항체 | |
BR112016013338B1 (pt) | Anticorpo pd-1 ou fragmento de ligação a antígeno do mesmo, seus usos e composição farmacêutica | |
CN107660150A (zh) | Il‑18结合蛋白(il‑18bp)和抗体在炎性疾病中 | |
CN105037541A (zh) | 具有高阻断活性的抗-C5a结合部分 | |
US20110311517A1 (en) | Antibodies and methods for treating estrogen receptor-associated diseases | |
JP7517999B2 (ja) | 抗IL-23p19抗体およびその使用 | |
CN109180816B (zh) | 抗人tim-3抗体及其用途 | |
BR112020013475A2 (pt) | Anticorpo de pd-l1, fragmento de ligação ao antígeno do mesmo, e uso farmacêutico do mesmo | |
CN105263964A (zh) | 抗-TNF-α/CXCL10双靶向抗体及其应用 | |
TW200825103A (en) | Novel polypeptides and uses thereof | |
CN113151186B (zh) | 抗人cd271的单克隆抗体及用途 | |
CN115066435A (zh) | 针对lilrb2的单域抗体 | |
TW202132338A (zh) | 針對類-tnf配體1a (tl1a)之人類化抗體及其用途 | |
CN103957938A (zh) | 治疗多发性骨髓瘤相关障碍的抗icam-1抗体 | |
CN108148133A (zh) | 抗vegf抗体 | |
WO2022044573A1 (ja) | コロナウイルススパイク蛋白に対するヒト抗体またはその抗原結合断片 | |
CN117264066B (zh) | 一种eno1抗体及其应用 | |
US9914768B2 (en) | Anti-S100A7 antibodies for the treatment and diagnosis of cancer | |
CN111944052A (zh) | 抗TNF-α/PD-1双特异性抗体及其应用 | |
WO2023109803A1 (zh) | 抗抑制素抗体及其应用 | |
CN109651509B (zh) | 抗cd20的人源化单抗及其制剂 | |
CN116333150A (zh) | 一种eno1抗体及其在治疗肿瘤中的应用 | |
CN114591424B (zh) | 新冠病毒s蛋白ntd区域的特异性抗体及其制备方法与应用 | |
WO2021160153A1 (zh) | 一种靶向人cd47的单域抗体及其用途 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |