WO2015147017A1 - Fgf2に対するアプタマー及びその使用 - Google Patents
Fgf2に対するアプタマー及びその使用 Download PDFInfo
- Publication number
- WO2015147017A1 WO2015147017A1 PCT/JP2015/058992 JP2015058992W WO2015147017A1 WO 2015147017 A1 WO2015147017 A1 WO 2015147017A1 JP 2015058992 W JP2015058992 W JP 2015058992W WO 2015147017 A1 WO2015147017 A1 WO 2015147017A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- aptamer
- fgf2
- nucleotide
- group
- ribose
- Prior art date
Links
- 108091023037 Aptamer Proteins 0.000 title claims abstract description 353
- 101150019331 FGF2 gene Proteins 0.000 title 1
- YSFTYXKQUONNFY-NQXPEFQPSA-N fgf2 Chemical compound C([C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](C)NC(=O)CNC(=O)[C@H](CO)NC(=O)CNC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H]1NCCC1)C1=CC=CC=C1 YSFTYXKQUONNFY-NQXPEFQPSA-N 0.000 title 1
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 claims abstract description 151
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 claims abstract description 149
- 239000003814 drug Substances 0.000 claims abstract description 34
- 239000000126 substance Substances 0.000 claims abstract description 26
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 12
- 229940079593 drug Drugs 0.000 claims abstract description 12
- 238000002372 labelling Methods 0.000 claims abstract description 10
- 238000012377 drug delivery Methods 0.000 claims abstract description 9
- 102000004190 Enzymes Human genes 0.000 claims abstract description 7
- 108090000790 Enzymes Proteins 0.000 claims abstract description 7
- 125000003729 nucleotide group Chemical group 0.000 claims description 124
- 239000002773 nucleotide Substances 0.000 claims description 123
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 claims description 60
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 claims description 60
- 230000027455 binding Effects 0.000 claims description 53
- 238000000034 method Methods 0.000 claims description 50
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 41
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 claims description 39
- 125000001153 fluoro group Chemical group F* 0.000 claims description 38
- 229910052731 fluorine Inorganic materials 0.000 claims description 36
- 125000004429 atom Chemical group 0.000 claims description 31
- 150000003230 pyrimidines Chemical class 0.000 claims description 26
- 239000002719 pyrimidine nucleotide Substances 0.000 claims description 25
- 108091008794 FGF receptors Proteins 0.000 claims description 24
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 24
- 230000033115 angiogenesis Effects 0.000 claims description 21
- 239000002213 purine nucleotide Substances 0.000 claims description 20
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 claims description 19
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 claims description 18
- 201000010099 disease Diseases 0.000 claims description 18
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 18
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 claims description 16
- 102000044168 Fibroblast Growth Factor Receptor Human genes 0.000 claims description 14
- 210000000988 bone and bone Anatomy 0.000 claims description 14
- 208000020084 Bone disease Diseases 0.000 claims description 11
- 208000015100 cartilage disease Diseases 0.000 claims description 11
- 230000036407 pain Effects 0.000 claims description 11
- 229910052799 carbon Inorganic materials 0.000 claims description 10
- 229940035893 uracil Drugs 0.000 claims description 9
- 229940113082 thymine Drugs 0.000 claims description 8
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 7
- 125000000561 purinyl group Chemical class N1=C(N=C2N=CNC2=C1)* 0.000 claims 4
- 230000006806 disease prevention Effects 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 27
- 230000002401 inhibitory effect Effects 0.000 abstract description 22
- 229920001223 polyethylene glycol Polymers 0.000 description 27
- 108020004707 nucleic acids Proteins 0.000 description 26
- 102000039446 nucleic acids Human genes 0.000 description 26
- 238000012986 modification Methods 0.000 description 25
- 230000004048 modification Effects 0.000 description 25
- 150000007523 nucleic acids Chemical class 0.000 description 25
- 239000002585 base Substances 0.000 description 24
- 108020003175 receptors Proteins 0.000 description 21
- 125000000548 ribosyl group Chemical group C1([C@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 21
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 20
- 125000005647 linker group Chemical group 0.000 description 20
- 102000005962 receptors Human genes 0.000 description 20
- -1 VEGF Proteins 0.000 description 19
- 102000004169 proteins and genes Human genes 0.000 description 18
- 108090000623 proteins and genes Proteins 0.000 description 18
- 150000001413 amino acids Chemical class 0.000 description 17
- 210000004027 cell Anatomy 0.000 description 16
- 150000003212 purines Chemical class 0.000 description 16
- 239000013076 target substance Substances 0.000 description 15
- 239000000243 solution Substances 0.000 description 14
- 108020004414 DNA Proteins 0.000 description 12
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 11
- 102000003971 Fibroblast Growth Factor 1 Human genes 0.000 description 11
- 108090000386 Fibroblast Growth Factor 1 Proteins 0.000 description 11
- 239000007790 solid phase Substances 0.000 description 11
- 102000052178 fibroblast growth factor receptor activity proteins Human genes 0.000 description 10
- 241001662443 Phemeranthus parviflorus Species 0.000 description 9
- 125000000524 functional group Chemical group 0.000 description 9
- 238000004519 manufacturing process Methods 0.000 description 9
- 238000000746 purification Methods 0.000 description 9
- 235000000346 sugar Nutrition 0.000 description 9
- 238000003786 synthesis reaction Methods 0.000 description 9
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 8
- 101100281008 Homo sapiens FGF2 gene Proteins 0.000 description 8
- 206010028980 Neoplasm Diseases 0.000 description 8
- 108091028043 Nucleic acid sequence Proteins 0.000 description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 8
- 239000011347 resin Substances 0.000 description 8
- 229920005989 resin Polymers 0.000 description 8
- 239000000523 sample Substances 0.000 description 8
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 7
- 239000004480 active ingredient Substances 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 230000006870 function Effects 0.000 description 7
- 239000003102 growth factor Substances 0.000 description 7
- 108090000765 processed proteins & peptides Proteins 0.000 description 7
- 239000000758 substrate Substances 0.000 description 7
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 6
- 108091008103 RNA aptamers Proteins 0.000 description 6
- 125000003277 amino group Chemical group 0.000 description 6
- 230000010261 cell growth Effects 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 6
- 229960002897 heparin Drugs 0.000 description 6
- 210000002997 osteoclast Anatomy 0.000 description 6
- 230000002265 prevention Effects 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 210000003556 vascular endothelial cell Anatomy 0.000 description 6
- 101800003838 Epidermal growth factor Proteins 0.000 description 5
- 102400001368 Epidermal growth factor Human genes 0.000 description 5
- 102000008108 Osteoprotegerin Human genes 0.000 description 5
- 108010035042 Osteoprotegerin Proteins 0.000 description 5
- 108010029485 Protein Isoforms Proteins 0.000 description 5
- 102000001708 Protein Isoforms Human genes 0.000 description 5
- 238000007792 addition Methods 0.000 description 5
- 239000000032 diagnostic agent Substances 0.000 description 5
- 229940039227 diagnostic agent Drugs 0.000 description 5
- 229940088598 enzyme Drugs 0.000 description 5
- 229940116977 epidermal growth factor Drugs 0.000 description 5
- 229920000669 heparin Polymers 0.000 description 5
- 239000003446 ligand Substances 0.000 description 5
- 108010082117 matrigel Proteins 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 239000000203 mixture Chemical class 0.000 description 5
- XXUPLYBCNPLTIW-UHFFFAOYSA-N octadec-7-ynoic acid Chemical compound CCCCCCCCCCC#CCCCCCC(O)=O XXUPLYBCNPLTIW-UHFFFAOYSA-N 0.000 description 5
- 239000002674 ointment Substances 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 238000011002 quantification Methods 0.000 description 5
- 206010039073 rheumatoid arthritis Diseases 0.000 description 5
- 238000006467 substitution reaction Methods 0.000 description 5
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 5
- 239000004094 surface-active agent Substances 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 5
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 4
- 102000003745 Hepatocyte Growth Factor Human genes 0.000 description 4
- 108090000100 Hepatocyte Growth Factor Proteins 0.000 description 4
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 239000002202 Polyethylene glycol Substances 0.000 description 4
- 229920001213 Polysorbate 20 Polymers 0.000 description 4
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 4
- 229920002472 Starch Polymers 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 239000011616 biotin Substances 0.000 description 4
- 229960002685 biotin Drugs 0.000 description 4
- 235000020958 biotin Nutrition 0.000 description 4
- 238000005859 coupling reaction Methods 0.000 description 4
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 4
- 239000003921 oil Substances 0.000 description 4
- 235000019198 oils Nutrition 0.000 description 4
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 4
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 4
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 230000001737 promoting effect Effects 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 235000019698 starch Nutrition 0.000 description 4
- 239000008107 starch Substances 0.000 description 4
- 238000013518 transcription Methods 0.000 description 4
- 230000035897 transcription Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 102100023995 Beta-nerve growth factor Human genes 0.000 description 3
- ZEOWTGPWHLSLOG-UHFFFAOYSA-N Cc1ccc(cc1-c1ccc2c(n[nH]c2c1)-c1cnn(c1)C1CC1)C(=O)Nc1cccc(c1)C(F)(F)F Chemical compound Cc1ccc(cc1-c1ccc2c(n[nH]c2c1)-c1cnn(c1)C1CC1)C(=O)Nc1cccc(c1)C(F)(F)F ZEOWTGPWHLSLOG-UHFFFAOYSA-N 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 3
- 101710182386 Fibroblast growth factor receptor 1 Proteins 0.000 description 3
- 102100023593 Fibroblast growth factor receptor 1 Human genes 0.000 description 3
- 229920002971 Heparan sulfate Polymers 0.000 description 3
- 101710163270 Nuclease Proteins 0.000 description 3
- 229920002684 Sepharose Polymers 0.000 description 3
- 239000007983 Tris buffer Substances 0.000 description 3
- 230000002159 abnormal effect Effects 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 206010064930 age-related macular degeneration Diseases 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 230000008878 coupling Effects 0.000 description 3
- 238000010168 coupling process Methods 0.000 description 3
- 238000012217 deletion Methods 0.000 description 3
- 230000037430 deletion Effects 0.000 description 3
- 230000004069 differentiation Effects 0.000 description 3
- 239000003937 drug carrier Substances 0.000 description 3
- 239000003480 eluent Substances 0.000 description 3
- 238000003780 insertion Methods 0.000 description 3
- 230000037431 insertion Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 208000002780 macular degeneration Diseases 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 235000019271 petrolatum Nutrition 0.000 description 3
- 108091033319 polynucleotide Proteins 0.000 description 3
- 239000002157 polynucleotide Substances 0.000 description 3
- 102000040430 polynucleotide Human genes 0.000 description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 3
- 229920000053 polysorbate 80 Polymers 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 239000003053 toxin Substances 0.000 description 3
- 231100000765 toxin Toxicity 0.000 description 3
- 108700012359 toxins Proteins 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- 210000003606 umbilical vein Anatomy 0.000 description 3
- JNYAEWCLZODPBN-JGWLITMVSA-N (2r,3r,4s)-2-[(1r)-1,2-dihydroxyethyl]oxolane-3,4-diol Chemical compound OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O JNYAEWCLZODPBN-JGWLITMVSA-N 0.000 description 2
- LVGUZGTVOIAKKC-UHFFFAOYSA-N 1,1,1,2-tetrafluoroethane Chemical compound FCC(F)(F)F LVGUZGTVOIAKKC-UHFFFAOYSA-N 0.000 description 2
- MYRTYDVEIRVNKP-UHFFFAOYSA-N 1,2-Divinylbenzene Chemical compound C=CC1=CC=CC=C1C=C MYRTYDVEIRVNKP-UHFFFAOYSA-N 0.000 description 2
- VBICKXHEKHSIBG-UHFFFAOYSA-N 1-monostearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(O)CO VBICKXHEKHSIBG-UHFFFAOYSA-N 0.000 description 2
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 2
- WLCZTRVUXYALDD-IBGZPJMESA-N 7-[[(2s)-2,6-bis(2-methoxyethoxycarbonylamino)hexanoyl]amino]heptoxy-methylphosphinic acid Chemical compound COCCOC(=O)NCCCC[C@H](NC(=O)OCCOC)C(=O)NCCCCCCCOP(C)(O)=O WLCZTRVUXYALDD-IBGZPJMESA-N 0.000 description 2
- 229930024421 Adenine Natural products 0.000 description 2
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 2
- 101710129634 Beta-nerve growth factor Proteins 0.000 description 2
- 208000006386 Bone Resorption Diseases 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 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 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 2
- 101150021185 FGF gene Proteins 0.000 description 2
- 102100031734 Fibroblast growth factor 19 Human genes 0.000 description 2
- 102100023600 Fibroblast growth factor receptor 2 Human genes 0.000 description 2
- 101710182389 Fibroblast growth factor receptor 2 Proteins 0.000 description 2
- 102100027844 Fibroblast growth factor receptor 4 Human genes 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 102000001554 Hemoglobins Human genes 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 2
- 101000846394 Homo sapiens Fibroblast growth factor 19 Proteins 0.000 description 2
- 101001052035 Homo sapiens Fibroblast growth factor 2 Proteins 0.000 description 2
- 101000917134 Homo sapiens Fibroblast growth factor receptor 4 Proteins 0.000 description 2
- 101000692455 Homo sapiens Platelet-derived growth factor receptor beta Proteins 0.000 description 2
- 101000808011 Homo sapiens Vascular endothelial growth factor A Proteins 0.000 description 2
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 2
- 206010023203 Joint destruction Diseases 0.000 description 2
- 239000004166 Lanolin Substances 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 102000005741 Metalloproteases Human genes 0.000 description 2
- 108010006035 Metalloproteases Proteins 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 2
- 239000004264 Petrolatum Substances 0.000 description 2
- 102100026547 Platelet-derived growth factor receptor beta Human genes 0.000 description 2
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 2
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 2
- 102100033810 RAC-alpha serine/threonine-protein kinase Human genes 0.000 description 2
- 102000014128 RANK Ligand Human genes 0.000 description 2
- 108010025832 RANK Ligand Proteins 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- PRXRUNOAOLTIEF-ADSICKODSA-N Sorbitan trioleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@@H](OC(=O)CCCCCCC\C=C/CCCCCCCC)[C@H]1OC[C@H](O)[C@H]1OC(=O)CCCCCCC\C=C/CCCCCCCC PRXRUNOAOLTIEF-ADSICKODSA-N 0.000 description 2
- 108010090804 Streptavidin Proteins 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 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 2
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 2
- DRTQHJPVMGBUCF-XVFCMESISA-N Uridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-XVFCMESISA-N 0.000 description 2
- 229960000643 adenine Drugs 0.000 description 2
- 238000005937 allylation reaction Methods 0.000 description 2
- 239000012491 analyte Substances 0.000 description 2
- 230000002491 angiogenic effect Effects 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 230000024279 bone resorption Effects 0.000 description 2
- 239000001506 calcium phosphate Substances 0.000 description 2
- 229910000389 calcium phosphate Inorganic materials 0.000 description 2
- 235000011010 calcium phosphates Nutrition 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 210000004748 cultured cell Anatomy 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 229940104302 cytosine Drugs 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 239000003405 delayed action preparation Substances 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 239000002612 dispersion medium Substances 0.000 description 2
- 210000002889 endothelial cell Anatomy 0.000 description 2
- 238000006200 ethylation reaction Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 210000002744 extracellular matrix Anatomy 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- BXWNKGSJHAJOGX-UHFFFAOYSA-N hexadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCO BXWNKGSJHAJOGX-UHFFFAOYSA-N 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 229920003063 hydroxymethyl cellulose Polymers 0.000 description 2
- 229940031574 hydroxymethyl cellulose Drugs 0.000 description 2
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 2
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 2
- 238000003018 immunoassay Methods 0.000 description 2
- 230000001976 improved effect Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 230000031146 intracellular signal transduction Effects 0.000 description 2
- NNPPMTNAJDCUHE-UHFFFAOYSA-N isobutane Chemical compound CC(C)C NNPPMTNAJDCUHE-UHFFFAOYSA-N 0.000 description 2
- 235000019388 lanolin Nutrition 0.000 description 2
- 229940039717 lanolin Drugs 0.000 description 2
- 235000010445 lecithin Nutrition 0.000 description 2
- 239000000787 lecithin Substances 0.000 description 2
- 229940067606 lecithin Drugs 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 229940092110 macugen Drugs 0.000 description 2
- 229910001629 magnesium chloride Inorganic materials 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
- 239000011159 matrix material Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 229920000609 methyl cellulose Polymers 0.000 description 2
- 238000007069 methylation reaction Methods 0.000 description 2
- 239000001923 methylcellulose Substances 0.000 description 2
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 2
- 238000013508 migration Methods 0.000 description 2
- 230000005012 migration Effects 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- ZIUHHBKFKCYYJD-UHFFFAOYSA-N n,n'-methylenebisacrylamide Chemical compound C=CC(=O)NCNC(=O)C=C ZIUHHBKFKCYYJD-UHFFFAOYSA-N 0.000 description 2
- 230000003472 neutralizing effect Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- GLDOVTGHNKAZLK-UHFFFAOYSA-N octadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCCCO GLDOVTGHNKAZLK-UHFFFAOYSA-N 0.000 description 2
- 125000001117 oleyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])/C([H])=C([H])\C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- 235000015205 orange juice Nutrition 0.000 description 2
- 210000000963 osteoblast Anatomy 0.000 description 2
- 125000004430 oxygen atom Chemical group O* 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 229940066842 petrolatum Drugs 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 229920001983 poloxamer Polymers 0.000 description 2
- 229920001296 polysiloxane Polymers 0.000 description 2
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 2
- 239000004810 polytetrafluoroethylene Substances 0.000 description 2
- 239000001103 potassium chloride Substances 0.000 description 2
- 235000011164 potassium chloride Nutrition 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 239000003380 propellant Substances 0.000 description 2
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 2
- ARIWANIATODDMH-UHFFFAOYSA-N rac-1-monolauroylglycerol Chemical compound CCCCCCCCCCCC(=O)OCC(O)CO ARIWANIATODDMH-UHFFFAOYSA-N 0.000 description 2
- 238000003127 radioimmunoassay Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 239000012146 running buffer Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 238000004904 shortening Methods 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 150000003431 steroids Chemical class 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 239000000454 talc Substances 0.000 description 2
- 229910052623 talc Inorganic materials 0.000 description 2
- 229940124597 therapeutic agent Drugs 0.000 description 2
- WYWHKKSPHMUBEB-UHFFFAOYSA-N tioguanine Chemical compound N1C(N)=NC(=S)C2=C1N=CN2 WYWHKKSPHMUBEB-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 239000003981 vehicle Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 230000029663 wound healing Effects 0.000 description 2
- NOOLISFMXDJSKH-UTLUCORTSA-N (+)-Neomenthol Chemical compound CC(C)[C@@H]1CC[C@@H](C)C[C@@H]1O NOOLISFMXDJSKH-UTLUCORTSA-N 0.000 description 1
- FPVKHBSQESCIEP-UHFFFAOYSA-N (8S)-3-(2-deoxy-beta-D-erythro-pentofuranosyl)-3,6,7,8-tetrahydroimidazo[4,5-d][1,3]diazepin-8-ol Natural products C1C(O)C(CO)OC1N1C(NC=NCC2O)=C2N=C1 FPVKHBSQESCIEP-UHFFFAOYSA-N 0.000 description 1
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 1
- FDKXTQMXEQVLRF-ZHACJKMWSA-N (E)-dacarbazine Chemical compound CN(C)\N=N\c1[nH]cnc1C(N)=O FDKXTQMXEQVLRF-ZHACJKMWSA-N 0.000 description 1
- LKJPYSCBVHEWIU-KRWDZBQOSA-N (R)-bicalutamide Chemical compound C([C@@](O)(C)C(=O)NC=1C=C(C(C#N)=CC=1)C(F)(F)F)S(=O)(=O)C1=CC=C(F)C=C1 LKJPYSCBVHEWIU-KRWDZBQOSA-N 0.000 description 1
- AJDIZQLSFPQPEY-UHFFFAOYSA-N 1,1,2-Trichlorotrifluoroethane Chemical compound FC(F)(Cl)C(F)(Cl)Cl AJDIZQLSFPQPEY-UHFFFAOYSA-N 0.000 description 1
- ZORQXIQZAOLNGE-UHFFFAOYSA-N 1,1-difluorocyclohexane Chemical compound FC1(F)CCCCC1 ZORQXIQZAOLNGE-UHFFFAOYSA-N 0.000 description 1
- DDMOUSALMHHKOS-UHFFFAOYSA-N 1,2-dichloro-1,1,2,2-tetrafluoroethane Chemical compound FC(F)(Cl)C(F)(F)Cl DDMOUSALMHHKOS-UHFFFAOYSA-N 0.000 description 1
- UHDGCWIWMRVCDJ-UHFFFAOYSA-N 1-beta-D-Xylofuranosyl-NH-Cytosine Natural products O=C1N=C(N)C=CN1C1C(O)C(O)C(CO)O1 UHDGCWIWMRVCDJ-UHFFFAOYSA-N 0.000 description 1
- OHMHBGPWCHTMQE-UHFFFAOYSA-N 2,2-dichloro-1,1,1-trifluoroethane Chemical compound FC(F)(F)C(Cl)Cl OHMHBGPWCHTMQE-UHFFFAOYSA-N 0.000 description 1
- JJCCTHHSPPINEI-MQWKRIRWSA-N 2,3-dihydroxypropyl (2s)-2,6-diaminohexanoate Chemical compound NCCCC[C@H](N)C(=O)OCC(O)CO JJCCTHHSPPINEI-MQWKRIRWSA-N 0.000 description 1
- 239000000263 2,3-dihydroxypropyl (Z)-octadec-9-enoate Substances 0.000 description 1
- HJRDNARELSKHEF-CLFAGFIQSA-N 2-[2-[(z)-octadec-9-enoyl]oxyethoxy]ethyl (z)-octadec-9-enoate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCCOCCOC(=O)CCCCCCC\C=C/CCCCCCCC HJRDNARELSKHEF-CLFAGFIQSA-N 0.000 description 1
- ASJSAQIRZKANQN-CRCLSJGQSA-N 2-deoxy-D-ribose Chemical group OC[C@@H](O)[C@@H](O)CC=O ASJSAQIRZKANQN-CRCLSJGQSA-N 0.000 description 1
- PMUNIMVZCACZBB-UHFFFAOYSA-N 2-hydroxyethylazanium;chloride Chemical compound Cl.NCCO PMUNIMVZCACZBB-UHFFFAOYSA-N 0.000 description 1
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 1
- RZRNAYUHWVFMIP-GDCKJWNLSA-N 3-oleoyl-sn-glycerol Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](O)CO RZRNAYUHWVFMIP-GDCKJWNLSA-N 0.000 description 1
- AOJJSUZBOXZQNB-VTZDEGQISA-N 4'-epidoxorubicin 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-VTZDEGQISA-N 0.000 description 1
- ZLOIGESWDJYCTF-UHFFFAOYSA-N 4-Thiouridine Natural products OC1C(O)C(CO)OC1N1C(=O)NC(=S)C=C1 ZLOIGESWDJYCTF-UHFFFAOYSA-N 0.000 description 1
- HIQIXEFWDLTDED-UHFFFAOYSA-N 4-hydroxy-1-piperidin-4-ylpyrrolidin-2-one Chemical compound O=C1CC(O)CN1C1CCNCC1 HIQIXEFWDLTDED-UHFFFAOYSA-N 0.000 description 1
- ZLOIGESWDJYCTF-XVFCMESISA-N 4-thiouridine Chemical group O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=S)C=C1 ZLOIGESWDJYCTF-XVFCMESISA-N 0.000 description 1
- LQLQRFGHAALLLE-UHFFFAOYSA-N 5-bromouracil Chemical group BrC1=CNC(=O)NC1=O LQLQRFGHAALLLE-UHFFFAOYSA-N 0.000 description 1
- KSNXJLQDQOIRIP-UHFFFAOYSA-N 5-iodouracil Chemical group IC1=CNC(=O)NC1=O KSNXJLQDQOIRIP-UHFFFAOYSA-N 0.000 description 1
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 1
- 244000215068 Acacia senegal Species 0.000 description 1
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 1
- 229920002126 Acrylic acid copolymer Polymers 0.000 description 1
- 229910002012 Aerosil® Inorganic materials 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 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
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 102100022987 Angiogenin Human genes 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 1
- 206010071155 Autoimmune arthritis Diseases 0.000 description 1
- 108010006654 Bleomycin Proteins 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 238000011746 C57BL/6J (JAX™ mouse strain) Methods 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- DLGOEMSEDOSKAD-UHFFFAOYSA-N Carmustine Chemical compound ClCCNC(=O)N(N=O)CCCl DLGOEMSEDOSKAD-UHFFFAOYSA-N 0.000 description 1
- 241000700198 Cavia Species 0.000 description 1
- 229920000623 Cellulose acetate phthalate Polymers 0.000 description 1
- 229920003043 Cellulose fiber Polymers 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- VOPWNXZWBYDODV-UHFFFAOYSA-N Chlorodifluoromethane Chemical compound FC(F)Cl VOPWNXZWBYDODV-UHFFFAOYSA-N 0.000 description 1
- PTOAARAWEBMLNO-KVQBGUIXSA-N Cladribine Chemical compound C1=NC=2C(N)=NC(Cl)=NC=2N1[C@H]1C[C@H](O)[C@@H](CO)O1 PTOAARAWEBMLNO-KVQBGUIXSA-N 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 1
- UHDGCWIWMRVCDJ-PSQAKQOGSA-N Cytidine Natural products O=C1N=C(N)C=CN1[C@@H]1[C@@H](O)[C@@H](O)[C@H](CO)O1 UHDGCWIWMRVCDJ-PSQAKQOGSA-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
- NOOLISFMXDJSKH-UHFFFAOYSA-N DL-menthol Natural products CC(C)C1CCC(C)CC1O NOOLISFMXDJSKH-UHFFFAOYSA-N 0.000 description 1
- 238000000018 DNA microarray Methods 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 101100015729 Drosophila melanogaster drk gene Proteins 0.000 description 1
- LVGKNOAMLMIIKO-UHFFFAOYSA-N Elaidinsaeure-aethylester Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC LVGKNOAMLMIIKO-UHFFFAOYSA-N 0.000 description 1
- 108010041308 Endothelial Growth Factors Proteins 0.000 description 1
- BRLQWZUYTZBJKN-UHFFFAOYSA-N Epichlorohydrin Chemical compound ClCC1CO1 BRLQWZUYTZBJKN-UHFFFAOYSA-N 0.000 description 1
- HTIJFSOGRVMCQR-UHFFFAOYSA-N Epirubicin Natural products COc1cccc2C(=O)c3c(O)c4CC(O)(CC(OC5CC(N)C(=O)C(C)O5)c4c(O)c3C(=O)c12)C(=O)CO HTIJFSOGRVMCQR-UHFFFAOYSA-N 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 239000001856 Ethyl cellulose Substances 0.000 description 1
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 description 1
- 229920003134 Eudragit® polymer Polymers 0.000 description 1
- 108010014173 Factor X Proteins 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 102100024802 Fibroblast growth factor 23 Human genes 0.000 description 1
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 1
- 206010017076 Fracture Diseases 0.000 description 1
- 208000010412 Glaucoma Diseases 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
- 108010024636 Glutathione Proteins 0.000 description 1
- IKAIKUBBJHFNBZ-LURJTMIESA-N Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CN IKAIKUBBJHFNBZ-LURJTMIESA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 102100033067 Growth factor receptor-bound protein 2 Human genes 0.000 description 1
- 108091009389 Growth factor receptor-bound protein 2 Proteins 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101001111439 Homo sapiens Beta-nerve growth factor Proteins 0.000 description 1
- 101100120051 Homo sapiens FGF1 gene Proteins 0.000 description 1
- 101001051973 Homo sapiens Fibroblast growth factor 23 Proteins 0.000 description 1
- 101000779418 Homo sapiens RAC-alpha serine/threonine-protein kinase Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- XDXDZDZNSLXDNA-TZNDIEGXSA-N Idarubicin Chemical compound C1[C@H](N)[C@H](O)[C@H](C)O[C@H]1O[C@@H]1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2C[C@@](O)(C(C)=O)C1 XDXDZDZNSLXDNA-TZNDIEGXSA-N 0.000 description 1
- XDXDZDZNSLXDNA-UHFFFAOYSA-N Idarubicin Natural products C1C(N)C(O)C(C)OC1OC1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2CC(O)(C(C)=O)C1 XDXDZDZNSLXDNA-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 208000008839 Kidney Neoplasms Diseases 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
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 102000043136 MAP kinase family Human genes 0.000 description 1
- 108091054455 MAP kinase family Proteins 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- CERQOIWHTDAKMF-UHFFFAOYSA-N Methacrylic acid Chemical compound CC(=C)C(O)=O CERQOIWHTDAKMF-UHFFFAOYSA-N 0.000 description 1
- VVQNEPGJFQJSBK-UHFFFAOYSA-N Methyl methacrylate Chemical compound COC(=O)C(C)=C VVQNEPGJFQJSBK-UHFFFAOYSA-N 0.000 description 1
- 229930192392 Mitomycin Natural products 0.000 description 1
- 101100390675 Mus musculus Fgf15 gene Proteins 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
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 206010029113 Neovascularisation Diseases 0.000 description 1
- 238000010934 O-alkylation reaction Methods 0.000 description 1
- 239000005642 Oleic acid Substances 0.000 description 1
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 description 1
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 description 1
- GOOHAUXETOMSMM-UHFFFAOYSA-N Propylene oxide Chemical group CC1CO1 GOOHAUXETOMSMM-UHFFFAOYSA-N 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 108091008611 Protein Kinase B Proteins 0.000 description 1
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 1
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 1
- 201000004681 Psoriasis Diseases 0.000 description 1
- 206010038389 Renal cancer Diseases 0.000 description 1
- 108091028664 Ribonucleotide Proteins 0.000 description 1
- 108010039491 Ricin Proteins 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 206010039710 Scleroderma Diseases 0.000 description 1
- 108010017898 Shiga Toxins Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- NWGKJDSIEKMTRX-AAZCQSIUSA-N Sorbitan monooleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O NWGKJDSIEKMTRX-AAZCQSIUSA-N 0.000 description 1
- 239000004147 Sorbitan trioleate Substances 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical group [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 235000019486 Sunflower oil Nutrition 0.000 description 1
- 229940123237 Taxane Drugs 0.000 description 1
- BPEGJWRSRHCHSN-UHFFFAOYSA-N Temozolomide Chemical compound O=C1N(C)N=NC2=C(C(N)=O)N=CN21 BPEGJWRSRHCHSN-UHFFFAOYSA-N 0.000 description 1
- 244000299461 Theobroma cacao Species 0.000 description 1
- 235000005764 Theobroma cacao ssp. cacao Nutrition 0.000 description 1
- 235000005767 Theobroma cacao ssp. sphaerocarpum Nutrition 0.000 description 1
- FOCVUCIESVLUNU-UHFFFAOYSA-N Thiotepa Chemical compound C1CN1P(N1CC1)(=S)N1CC1 FOCVUCIESVLUNU-UHFFFAOYSA-N 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- IVTVGDXNLFLDRM-HNNXBMFYSA-N Tomudex Chemical compound C=1C=C2NC(C)=NC(=O)C2=CC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)S1 IVTVGDXNLFLDRM-HNNXBMFYSA-N 0.000 description 1
- 108020004566 Transfer RNA Proteins 0.000 description 1
- PHYFQTYBJUILEZ-UHFFFAOYSA-N Trioleoylglycerol Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC(OC(=O)CCCCCCCC=CCCCCCCCC)COC(=O)CCCCCCCC=CCCCCCCCC PHYFQTYBJUILEZ-UHFFFAOYSA-N 0.000 description 1
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 1
- 108091008605 VEGF receptors Proteins 0.000 description 1
- 108010073923 Vascular Endothelial Growth Factor C Proteins 0.000 description 1
- 102000009520 Vascular Endothelial Growth Factor C Human genes 0.000 description 1
- 102100033177 Vascular endothelial growth factor receptor 2 Human genes 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
- 229940122803 Vinca alkaloid Drugs 0.000 description 1
- 238000002441 X-ray diffraction Methods 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- IYKJEILNJZQJPU-UHFFFAOYSA-N acetic acid;butanedioic acid Chemical compound CC(O)=O.OC(=O)CCC(O)=O IYKJEILNJZQJPU-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 238000005804 alkylation reaction Methods 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- CEGOLXSVJUTHNZ-UHFFFAOYSA-K aluminium tristearate Chemical compound [Al+3].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O CEGOLXSVJUTHNZ-UHFFFAOYSA-K 0.000 description 1
- 229940063655 aluminum stearate Drugs 0.000 description 1
- 229940059260 amidate Drugs 0.000 description 1
- 238000005576 amination reaction Methods 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 108010072788 angiogenin Proteins 0.000 description 1
- 229940045799 anthracyclines and related substance Drugs 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000003432 anti-folate effect Effects 0.000 description 1
- 229940125644 antibody drug Drugs 0.000 description 1
- 229940127074 antifolate Drugs 0.000 description 1
- 229940045686 antimetabolites antineoplastic purine analogs Drugs 0.000 description 1
- 229940045713 antineoplastic alkylating drug ethylene imines Drugs 0.000 description 1
- 229940045688 antineoplastic antimetabolites pyrimidine analogues Drugs 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 229940045696 antineoplastic drug podophyllotoxin derivative Drugs 0.000 description 1
- 229940045985 antineoplastic platinum compound Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 229910052586 apatite Inorganic materials 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 108010045569 atelocollagen Proteins 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000003305 autocrine Effects 0.000 description 1
- 229940120638 avastin Drugs 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
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000037429 base substitution Effects 0.000 description 1
- 235000013871 bee wax Nutrition 0.000 description 1
- 239000012166 beeswax Substances 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
- DRTQHJPVMGBUCF-PSQAKQOGSA-N beta-L-uridine Natural products O[C@H]1[C@@H](O)[C@H](CO)O[C@@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-PSQAKQOGSA-N 0.000 description 1
- 229960000997 bicalutamide Drugs 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 102000023732 binding proteins Human genes 0.000 description 1
- 108091008324 binding proteins Proteins 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 229960001561 bleomycin Drugs 0.000 description 1
- 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 1
- 229920001400 block copolymer Polymers 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 235000014121 butter Nutrition 0.000 description 1
- 235000001046 cacaotero Nutrition 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- FNAQSUUGMSOBHW-UHFFFAOYSA-H calcium citrate Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FNAQSUUGMSOBHW-UHFFFAOYSA-H 0.000 description 1
- 239000001354 calcium citrate Substances 0.000 description 1
- HXCHCVDVKSCDHU-LULTVBGHSA-N calicheamicin Chemical compound C1[C@H](OC)[C@@H](NCC)CO[C@H]1O[C@H]1[C@H](O[C@@H]2C\3=C(NC(=O)OC)C(=O)C[C@](C/3=C/CSSSC)(O)C#C\C=C/C#C2)O[C@H](C)[C@@H](NO[C@@H]2O[C@H](C)[C@@H](SC(=O)C=3C(=C(OC)C(O[C@H]4[C@@H]([C@H](OC)[C@@H](O)[C@H](C)O4)O)=C(I)C=3C)OC)[C@@H](O)C2)[C@@H]1O HXCHCVDVKSCDHU-LULTVBGHSA-N 0.000 description 1
- 229930195731 calicheamicin Natural products 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 1
- 229960004562 carboplatin Drugs 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 229960005243 carmustine Drugs 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 235000019438 castor oil Nutrition 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 229940081734 cellulose acetate phthalate Drugs 0.000 description 1
- 229960000541 cetyl alcohol Drugs 0.000 description 1
- 229960001927 cetylpyridinium chloride Drugs 0.000 description 1
- YMKDRGPMQRFJGP-UHFFFAOYSA-M cetylpyridinium chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCC[N+]1=CC=CC=C1 YMKDRGPMQRFJGP-UHFFFAOYSA-M 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 229960004106 citric acid Drugs 0.000 description 1
- 229960002436 cladribine Drugs 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 239000002285 corn oil Substances 0.000 description 1
- 235000005687 corn oil Nutrition 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 239000002385 cottonseed oil Substances 0.000 description 1
- 229920006037 cross link polymer Polymers 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- YPHMISFOHDHNIV-FSZOTQKASA-N cycloheximide Chemical compound C1[C@@H](C)C[C@H](C)C(=O)[C@@H]1[C@H](O)CC1CC(=O)NC(=O)C1 YPHMISFOHDHNIV-FSZOTQKASA-N 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- UHDGCWIWMRVCDJ-ZAKLUEHWSA-N cytidine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O1 UHDGCWIWMRVCDJ-ZAKLUEHWSA-N 0.000 description 1
- 229960003901 dacarbazine Drugs 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 229940124447 delivery agent Drugs 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- PXBRQCKWGAHEHS-UHFFFAOYSA-N dichlorodifluoromethane Chemical compound FC(F)(Cl)Cl PXBRQCKWGAHEHS-UHFFFAOYSA-N 0.000 description 1
- 235000019404 dichlorodifluoromethane Nutrition 0.000 description 1
- 229910001873 dinitrogen Inorganic materials 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 229960003668 docetaxel Drugs 0.000 description 1
- 125000003438 dodecyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 229960004679 doxorubicin Drugs 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- VQNATVDKACXKTF-XELLLNAOSA-N duocarmycin Chemical compound COC1=C(OC)C(OC)=C2NC(C(=O)N3C4=CC(=O)C5=C([C@@]64C[C@@H]6C3)C=C(N5)C(=O)OC)=CC2=C1 VQNATVDKACXKTF-XELLLNAOSA-N 0.000 description 1
- 229960005501 duocarmycin Drugs 0.000 description 1
- 229930184221 duocarmycin Natural products 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229960001904 epirubicin Drugs 0.000 description 1
- 229960001842 estramustine Drugs 0.000 description 1
- FRPJXPJMRWBBIH-RBRWEJTLSA-N estramustine Chemical compound ClCCN(CCCl)C(=O)OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 FRPJXPJMRWBBIH-RBRWEJTLSA-N 0.000 description 1
- HQPMKSGTIOYHJT-UHFFFAOYSA-N ethane-1,2-diol;propane-1,2-diol Chemical compound OCCO.CC(O)CO HQPMKSGTIOYHJT-UHFFFAOYSA-N 0.000 description 1
- 150000002170 ethers Chemical class 0.000 description 1
- 235000019325 ethyl cellulose Nutrition 0.000 description 1
- 229920001249 ethyl cellulose Polymers 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- LVGKNOAMLMIIKO-QXMHVHEDSA-N ethyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC LVGKNOAMLMIIKO-QXMHVHEDSA-N 0.000 description 1
- 229940093471 ethyl oleate Drugs 0.000 description 1
- NPUKDXXFDDZOKR-LLVKDONJSA-N etomidate Chemical compound CCOC(=O)C1=CN=CN1[C@H](C)C1=CC=CC=C1 NPUKDXXFDDZOKR-LLVKDONJSA-N 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
- 229960005420 etoposide Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 229960000390 fludarabine Drugs 0.000 description 1
- GIUYCYHIANZCFB-FJFJXFQQSA-N fludarabine phosphate Chemical compound C1=NC=2C(N)=NC(F)=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@@H]1O GIUYCYHIANZCFB-FJFJXFQQSA-N 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 102000034287 fluorescent proteins Human genes 0.000 description 1
- 108091006047 fluorescent proteins Proteins 0.000 description 1
- 238000003682 fluorination reaction Methods 0.000 description 1
- 229960002949 fluorouracil Drugs 0.000 description 1
- 239000004052 folic acid antagonist Substances 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 238000009472 formulation Methods 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
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 1
- 229940068939 glyceryl monolaurate Drugs 0.000 description 1
- 229940075507 glyceryl monostearate Drugs 0.000 description 1
- 108010015792 glycyllysine Proteins 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 101150098203 grb2 gene Proteins 0.000 description 1
- 210000003566 hemangioblast Anatomy 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 102000058223 human VEGFA Human genes 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000008311 hydrophilic ointment Substances 0.000 description 1
- 229920001477 hydrophilic polymer Polymers 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 239000001341 hydroxy propyl starch Substances 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 1
- 229920003132 hydroxypropyl methylcellulose phthalate Polymers 0.000 description 1
- 229940031704 hydroxypropyl methylcellulose phthalate Drugs 0.000 description 1
- 235000013828 hydroxypropyl starch Nutrition 0.000 description 1
- 229960000908 idarubicin Drugs 0.000 description 1
- 229960001101 ifosfamide Drugs 0.000 description 1
- HOMGKSMUEGBAAB-UHFFFAOYSA-N ifosfamide Chemical compound ClCCNP1(=O)OCCCN1CCCl HOMGKSMUEGBAAB-UHFFFAOYSA-N 0.000 description 1
- 239000012729 immediate-release (IR) formulation Substances 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011503 in vivo imaging Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 239000001282 iso-butane Substances 0.000 description 1
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 1
- 229940074928 isopropyl myristate Drugs 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 239000003350 kerosene Substances 0.000 description 1
- 201000010982 kidney cancer Diseases 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 125000000400 lauroyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 125000005645 linoleyl group Chemical group 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 229960003511 macrogol Drugs 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 229910001437 manganese ion Inorganic materials 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 230000000873 masking effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- HAWPXGHAZFHHAD-UHFFFAOYSA-N mechlorethamine Chemical class ClCCN(C)CCCl HAWPXGHAZFHHAD-UHFFFAOYSA-N 0.000 description 1
- 229960001924 melphalan Drugs 0.000 description 1
- 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 1
- 229940041616 menthol Drugs 0.000 description 1
- 229920003145 methacrylic acid copolymer Polymers 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 description 1
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 1
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 1
- 229960002216 methylparaben Drugs 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 229960003775 miltefosine Drugs 0.000 description 1
- PQLXHQMOHUQAKB-UHFFFAOYSA-N miltefosine Chemical compound CCCCCCCCCCCCCCCCOP([O-])(=O)OCC[N+](C)(C)C PQLXHQMOHUQAKB-UHFFFAOYSA-N 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- 229960001156 mitoxantrone Drugs 0.000 description 1
- KKZJGLLVHKMTCM-UHFFFAOYSA-N mitoxantrone Chemical compound O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO KKZJGLLVHKMTCM-UHFFFAOYSA-N 0.000 description 1
- 239000003607 modifier Substances 0.000 description 1
- 239000001788 mono and diglycerides of fatty acids Substances 0.000 description 1
- RZRNAYUHWVFMIP-UHFFFAOYSA-N monoelaidin Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC(O)CO RZRNAYUHWVFMIP-UHFFFAOYSA-N 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- IJDNQMDRQITEOD-UHFFFAOYSA-N n-butane Chemical compound CCCC IJDNQMDRQITEOD-UHFFFAOYSA-N 0.000 description 1
- GOQYKNQRPGWPLP-UHFFFAOYSA-N n-heptadecyl alcohol Natural products CCCCCCCCCCCCCCCCCO GOQYKNQRPGWPLP-UHFFFAOYSA-N 0.000 description 1
- 239000006199 nebulizer Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 201000003142 neovascular glaucoma Diseases 0.000 description 1
- OSTGTTZJOCZWJG-UHFFFAOYSA-N nitrosourea Chemical compound NC(=O)N=NO OSTGTTZJOCZWJG-UHFFFAOYSA-N 0.000 description 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 1
- 230000005937 nuclear translocation Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000000414 obstructive effect Effects 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 125000002811 oleoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])/C([H])=C([H])\C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 230000011164 ossification Effects 0.000 description 1
- 201000008482 osteoarthritis Diseases 0.000 description 1
- 229940045795 other cytotoxic antibiotic in ATC Drugs 0.000 description 1
- 229960001756 oxaliplatin Drugs 0.000 description 1
- DWAFYCQODLXJNR-BNTLRKBRSA-L oxaliplatin Chemical compound O1C(=O)C(=O)O[Pt]11N[C@@H]2CCCC[C@H]2N1 DWAFYCQODLXJNR-BNTLRKBRSA-L 0.000 description 1
- GIPDEPRRXIBGNF-KTKRTIGZSA-N oxolan-2-ylmethyl (z)-octadec-9-enoate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC1CCCO1 GIPDEPRRXIBGNF-KTKRTIGZSA-N 0.000 description 1
- 125000006353 oxyethylene group Chemical group 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 125000001312 palmitoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000003076 paracrine Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- VSIIXMUUUJUKCM-UHFFFAOYSA-D pentacalcium;fluoride;triphosphate Chemical compound [F-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O VSIIXMUUUJUKCM-UHFFFAOYSA-D 0.000 description 1
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 1
- 229960002340 pentostatin Drugs 0.000 description 1
- FPVKHBSQESCIEP-JQCXWYLXSA-N pentostatin Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(N=CNC[C@H]2O)=C2N=C1 FPVKHBSQESCIEP-JQCXWYLXSA-N 0.000 description 1
- 208000028169 periodontal disease Diseases 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 150000003058 platinum compounds Chemical class 0.000 description 1
- YJGVMLPVUAXIQN-XVVDYKMHSA-N podophyllotoxin Chemical class COC1=C(OC)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@H](O)[C@@H]3[C@@H]2C(OC3)=O)=C1 YJGVMLPVUAXIQN-XVVDYKMHSA-N 0.000 description 1
- 239000003600 podophyllotoxin derivative Substances 0.000 description 1
- 229920001993 poloxamer 188 Polymers 0.000 description 1
- 229920001606 poly(lactic acid-co-glycolic acid) Polymers 0.000 description 1
- 229940068918 polyethylene glycol 400 Drugs 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 159000000001 potassium salts Chemical class 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 239000001294 propane Substances 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 description 1
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 239000000941 radioactive substance Substances 0.000 description 1
- 229960004432 raltitrexed Drugs 0.000 description 1
- 108700015048 receptor decoy activity proteins Proteins 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000002336 ribonucleotide Substances 0.000 description 1
- 125000002652 ribonucleotide group Chemical group 0.000 description 1
- 238000003118 sandwich ELISA Methods 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 210000000329 smooth muscle myocyte Anatomy 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 229940083542 sodium Drugs 0.000 description 1
- 235000015424 sodium Nutrition 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 229940001607 sodium bisulfite Drugs 0.000 description 1
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 1
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 1
- 235000011069 sorbitan monooleate Nutrition 0.000 description 1
- 239000001593 sorbitan monooleate Substances 0.000 description 1
- 229940035049 sorbitan monooleate Drugs 0.000 description 1
- 235000019337 sorbitan trioleate Nutrition 0.000 description 1
- 229960000391 sorbitan trioleate Drugs 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 125000003696 stearoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 229940012831 stearyl alcohol Drugs 0.000 description 1
- 125000004079 stearyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 239000002600 sunflower oil Substances 0.000 description 1
- 230000008093 supporting effect Effects 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 210000002437 synoviocyte Anatomy 0.000 description 1
- 229920001059 synthetic polymer Polymers 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- DKPFODGZWDEEBT-QFIAKTPHSA-N taxane Chemical class C([C@]1(C)CCC[C@@H](C)[C@H]1C1)C[C@H]2[C@H](C)CC[C@@H]1C2(C)C DKPFODGZWDEEBT-QFIAKTPHSA-N 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
- 229960001674 tegafur Drugs 0.000 description 1
- WFWLQNSHRPWKFK-ZCFIWIBFSA-N tegafur Chemical compound O=C1NC(=O)C(F)=CN1[C@@H]1OCCC1 WFWLQNSHRPWKFK-ZCFIWIBFSA-N 0.000 description 1
- 229960004964 temozolomide Drugs 0.000 description 1
- 229940072958 tetrahydrofurfuryl oleate Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 229960001196 thiotepa Drugs 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 229960003087 tioguanine Drugs 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 239000004408 titanium dioxide Substances 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 229960000303 topotecan Drugs 0.000 description 1
- UCFGDBYHRUNTLO-QHCPKHFHSA-N topotecan Chemical compound C1=C(O)C(CN(C)C)=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 UCFGDBYHRUNTLO-QHCPKHFHSA-N 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 235000013337 tricalcium citrate Nutrition 0.000 description 1
- CYRMSUTZVYGINF-UHFFFAOYSA-N trichlorofluoromethane Chemical compound FC(Cl)(Cl)Cl CYRMSUTZVYGINF-UHFFFAOYSA-N 0.000 description 1
- PHYFQTYBJUILEZ-IUPFWZBJSA-N triolein Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(OC(=O)CCCCCCC\C=C/CCCCCCCC)COC(=O)CCCCCCC\C=C/CCCCCCCC PHYFQTYBJUILEZ-IUPFWZBJSA-N 0.000 description 1
- 229940117972 triolein Drugs 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 229960000875 trofosfamide Drugs 0.000 description 1
- UMKFEPPTGMDVMI-UHFFFAOYSA-N trofosfamide Chemical compound ClCCN(CCCl)P1(=O)OCCCN1CCCl UMKFEPPTGMDVMI-UHFFFAOYSA-N 0.000 description 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- DRTQHJPVMGBUCF-UHFFFAOYSA-N uracil arabinoside Natural products OC1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-UHFFFAOYSA-N 0.000 description 1
- 229940045145 uridine Drugs 0.000 description 1
- 208000019553 vascular disease Diseases 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 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
- 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
- 229960004528 vincristine Drugs 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
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- 239000003871 white petrolatum Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/712—Nucleic acids or oligonucleotides having modified sugars, i.e. other than ribose or 2'-deoxyribose
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/06—Antipsoriatics
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/04—Centrally acting analgesics, e.g. opioids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
- A61P27/06—Antiglaucoma agents or miotics
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
- C07H21/02—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
- C07H21/04—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/115—Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2121/00—Preparations for use in therapy
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/16—Aptamers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/31—Chemical structure of the backbone
- C12N2310/317—Chemical structure of the backbone with an inverted bond, e.g. a cap structure
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/32—Chemical structure of the sugar
- C12N2310/321—2'-O-R Modification
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/32—Chemical structure of the sugar
- C12N2310/322—2'-R Modification
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/34—Spatial arrangement of the modifications
- C12N2310/344—Position-specific modifications, e.g. on every purine, at the 3'-end
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/35—Nature of the modification
- C12N2310/352—Nature of the modification linked to the nucleic acid via a carbon atom
- C12N2310/3521—Methyl
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/35—Nature of the modification
- C12N2310/353—Nature of the modification linked to the nucleic acid via an atom other than carbon
- C12N2310/3533—Halogen
Definitions
- the present invention relates to an aptamer for FGF2 and a method for using the aptamer.
- FGF2 Basic fibroblast growth factor
- bFGF Basic fibroblast growth factor
- human FGF2 has a plurality of isoforms, only the isoform having the smallest molecular weight is secreted extracellularly.
- This isoform is a protein of about 18 kDa composed of 154 amino acids, has no sugar chain, and has an isoelectric point of 9.4, which is inclined basic.
- the function of high molecular weight isoforms (22, 22.5, 24, 34 kD) of FGF2 with different reading frames is not yet clear, but it has a nuclear translocation signal and is thought to function in the nucleus.
- FGF2 is classified into the FGF1 subfamily along with FGF1.
- the homology of the amino acid sequence with FGF1 is the highest among all FGFs, and its value is 55%.
- Receptors for FGF (FGFR) are tyrosine kinase type receptors and are classified into four subtypes.
- FGFR1-3 are known to have isoforms b and c, respectively.
- FGF2 binds to FGFR1b, FGFR1c, FGFR2c and FGFR3c, and FGFR4, and dimerizes these receptors.
- Mouse fibroblasts are cells expressing FGFR1 on the cell membrane surface. This FGFR1 is known to be activated by the binding of human FGF2.
- FGF2 binds to FGFR1
- MAP Kinase miratePitropin3
- FRS2 Fibroblast growth factor receptor substream 2
- Grb2 growth factor receptor-bound protein 2
- MAP kinase-mPI / AKT1 protein kinase B pathway
- VEGF basic endothelial growth factor
- VEGF-A hepatocyte growth factor
- HGF hepatocyte growth factor
- angry factor oietin-2 VEGFR
- PDGFR- ⁇ platelet-derived growth factor beta receptor- ⁇
- FGF2 has a heparin-binding region and binds to heparin and heparan sulfate like other FGFs. It is believed that FGF2 secreted from cells generally binds to heparan sulfate in the extracellular matrix, is concentrated, and is protected from proteases. When functioning as a ligand, release from the bound extracellular matrix is required, but it has been reported that FGF-BP (FGF-binding protein) is involved in this and assists the induction into FGFR.
- FGF-BP FGF-binding protein
- FGF2 has a strong proliferation and migration promoting action on vascular endothelial cells and is deeply involved in angiogenesis of tumor tissue.
- the serum concentration of FGF2 is particularly high in tumors with many blood vessels such as kidney cancer, and is also present in various tumors such as prostate cancer, breast cancer and lung cancer.
- vascularization includes FGF1, VEGF, TNF- ⁇ (tumor necrosis factor- ⁇ ), PDGF, EGF (epidermal growth factor), MMP (matrix metallopeptidase), and angiogenin.
- FGF1 vascular endothelial growth factor
- VEGF tumor necrosis factor- ⁇
- PDGF vascular endothelial growth factor
- EGF epidermal growth factor
- MMP matrix metallopeptidase
- angiogenin angiogenin.
- FGF2 differs from other factors in that it acts not only on vascular endothelial cells but also on mesenchymal cells surrounding endothelial cells such as smooth muscle cells. That is, FGF2 is considered to stimulate mesenchymal cells to enhance the expression of PDGF, PDGFR, VEGF, HGF, and the like, and these factors promote direct proliferation of vascular endothelial cells.
- abnormal angiogenesis is also involved in diseases such as periodontal disease, scleroderma, neovascular glaucoma, chronic inflammation such as arthritis, psoriasis, and age-related macular degeneration.
- FGF2 is known to have an effect of promoting bone formation, but on the other hand, it is also attracting attention as a factor for promoting bone resorption, such as being involved in joint destruction in patients with rheumatoid arthritis.
- rheumatoid arthritis characterized by autoimmune arthritis, osteoclasts increase, bone resorption increases, and bone destruction progresses.
- FGF2 stimulates mesenchymal cells to enhance the expression of inflammatory cytokines and growth factors such as PDGF, PDGFR, VEGF, and HGF, promote angiogenesis, and promote bone destruction. That is, it is known that FGF2 is a central molecule involved in an important disease state in rheumatoid arthritis (see Non-Patent Document 1).
- Osteoprotegerin is a decoy receptor for RANKL, an osteoclast-inducing factor, and is known to antagonize RANK and inhibit differentiation into osteoclasts and its function. (See Non-Patent Document 2). It is also known that OPG produced from synovial cells is suppressed by FGF2 stimulation (see Non-Patent Document 3). Furthermore, FGF2 promotes the coupling of osteoblasts and osteoclast precursor cells by inducing high expression of RANKL in osteoblasts, and consequently promotes differentiation and activation into osteoclasts (see Non-Patent Document 4). ).
- RNA aptamers are an RNA that specifically binds to a target substance such as a protein, and can be prepared using the SELEX method (Systematic Evolution of Keywords by Exponential Enrichment) (see Patent Documents 1 to 3).
- the SELEX method is a method of selecting RNA that specifically binds to a target substance from a pool of RNA having about 10 14 different nucleotide sequences.
- the RNA used has a structure in which a random sequence of about 40 residues is sandwiched between primer sequences. This RNA pool is associated with the target substance, and only the RNA bound to the target substance is recovered using a filter or the like. The recovered RNA is amplified by RT-PCR and used as a template for the next round. By repeating this operation about 10 times, an RNA aptamer that specifically binds to the target substance may be obtained.
- Patent Document 4 describes an aptamer that binds to FGF2 obtained by the SELEX method. However, the aptamer differs in sequence from the aptamer specifically shown herein. In addition, this document does not suggest any aptamer specifically shown in the present specification.
- An object of the present invention is to provide an aptamer for FGF2 and a method for using the aptamer.
- the present inventor succeeded in producing a high-quality aptamer for FGF2 and thus completed the present invention.
- N 1 and N 6 are each independently any 0 to several bases, N 2 , N 3 , N 4 and N 5 independently represent any one base, (A) a nucleotide contained in the aptamer, (I) the ribose 2 'position of each pyrimidine nucleotide is a fluorine atom; (Ii) an aptamer in which the ribose 2 ′ position of each purine nucleotide is a hydroxy group; (B) in the aptamer of (a), (I) The 2'-position fluorine atom of ribo
- N 1 is G, GG, AG, C or a gap
- N 2 is A or U
- N 3 is G, C or A
- N 4 is G, C or U
- N 5 is G or U
- N 6 is UUCN 61 or AGUCN 62
- N 61 and N 62 are each independently any 0 to several bases
- [3] The aptamer according to [1] or [2], comprising a nucleotide sequence represented by the following formula (2) or (3): GGGAAACUAGGGCGUUAACGUGACCAGGUGUUCN 61 (Formula 2)
- N 1 GGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCN 62 (Formula 3).
- aptamer according to any one of [1] to [5], wherein the aptamer includes a nucleotide sequence in which one or several nucleotides are substituted, deleted, inserted or added, (A) a nucleotide contained in the aptamer, (I) the ribose 2 'position of each pyrimidine nucleotide is a fluorine atom; (Ii) an aptamer in which the ribose 2 ′ position of each purine nucleotide is a hydroxy group; (B) in the aptamer of (a), (I) The 2'-position fluorine atom of ribose of each pyrimidine nucleotide is each independently unsubstituted or substituted with an atom or group selected from the group consisting of a hydrogen atom, a hydroxy group and a methoxy group , (Ii) The 2'-position hydroxy group of ribose of each purine
- [7] The aptamer according to any one of [1] to [6], wherein the nucleotide has a length of 45 nucleotides or less.
- [8] The aptamer according to any one of [1] to [7], which inhibits binding between FGF2 and an FGF receptor.
- [10] A complex comprising the aptamer according to any one of [1] to [9] and a functional substance.
- the functional substance is an affinity substance, a labeling substance, an enzyme, a drug delivery vehicle or a drug.
- a medicament comprising the aptamer according to any one of [1] to [9] or the complex according to [10] or [11].
- Treatment or prevention of a disease associated with angiogenesis, bone / cartilage disease or pain comprising the aptamer according to any one of [1] to [9] or the complex according to [10] or [11] Pharmaceuticals.
- a method for treating or preventing a disease associated with angiogenesis, a bone / cartilage disease or pain wherein the aptamer according to any one of [1] to [9] or [10] or [11]
- a method comprising administering to the subject the complex described in 1.
- the aptamer according to any one of [1] to [9] or [10] or [11] for producing a medicament for the treatment or prevention of a disease associated with angiogenesis, bone / cartilage disease or pain Use of the complex described in 1.
- the aptamer or complex of the present invention may be useful as a therapeutic or prophylactic agent for a disease associated with angiogenesis, a bone / cartilage disease or pain, or a diagnostic agent or reagent.
- the aptamers or complexes of the present invention may also be useful for FGF2 purification and enrichment, FGF2 labeling, and FGF2 detection and quantification.
- FIG. 1 shows a sensorgram showing that an aptamer represented by aptamer IDs 1 and 2 binds to human FGF2.
- FIG. 2 shows a sensorgram in which the aptamer represented by aptamer ID1 inhibits the binding of human FGF2 and four receptors.
- FIG. 3 shows a sensorgram indicating that the aptamer represented by aptamer ID3 does not bind to human FGF1, EGF, NGF, or VEGF.
- N 1 and N 6 are each independently any 0 to several bases, N 2 , N 3 , N 4 and N 5 independently represent any one base,
- An aptamer comprising a nucleotide sequence represented by (wherein uracil may be thymine), wherein (a) or (b) (A) a nucleotide contained in the aptamer, (I) the ribose 2 'position of each pyrimidine nucleotide is a fluorine atom; (Ii) an aptamer in which the 2 ′ position of ribose of each purine nucleotide is a hydroxy group; or (b) in the aptamer of (a), (I) The 2'-position fluorine atom of ribose of each pyrimidine nucleotide is each independently unsub
- Aptamer refers to a nucleic acid molecule having binding activity to a predetermined target molecule. Aptamers can inhibit the activity of a given target molecule by binding to the given target molecule.
- the aptamer of the present invention may be RNA, DNA, modified nucleic acid or a mixture thereof.
- the aptamer of the present invention may be in a linear or cyclic form.
- the present invention provides an aptamer having binding activity to FGF2.
- the aptamer of the invention can bind to FGF2 and inhibit the activity of FGF2. That is, the aptamer of the present invention may have an inhibitory activity against FGF2.
- the inhibitory activity against FGF2 means the ability to inhibit any activity possessed by FGF2.
- FGF2 acts on FGF receptor-expressing cells, activates signal transduction, and induces production of various cell growth factors and their receptors. Therefore, the inhibitory activity against FGF2 can be an activity that inhibits intracellular signal transduction via the FGF receptor.
- expression of these various cell growth factors and their receptors results in enhancement of cell growth activity and migration activity, and thus FGF2 inhibitory activity means inhibition of those activities.
- the aptamer of the present invention binds to FGF2 and inhibits the binding between FGF2 and the FGF receptor, the action accompanying the activation of the intracellular signal transduction pathway via the FGF receptor, for example, suppression of cell death, Cell proliferation, suppression of osteoprotegerin (OPG) production, and the like can be inhibited.
- FGF2 is a protein that is strongly expressed at the early stage of development, differentiation, proliferation, and regeneration.
- FGF2 is a protein having an amino acid sequence represented by Accession code EAX05222 or NP001997.
- FGF2 may also be called bFGF (basic FGF), FGFB, or HBGF-2.
- FGF2 in the present invention can be produced not only in the body of an animal but also in mammalian cells such as mice, insect cells, cultured cells such as Escherichia coli, and can also be produced by chemical synthesis. When producing by cultured cells or chemical synthesis, a mutant can be easily produced by a method known per se.
- the “mutant” of FGF2 is one in which one to several amino acids are substituted, deleted, added, or the like from the known FGF2 amino acid sequence, or a partial amino acid sequence of the known FGF2 amino acid sequence. It means a protein or peptide having at least one activity that FGF2 originally has. When an amino acid is substituted or added, the amino acid may be a natural amino acid or a non-natural amino acid. FGF2 in the present invention includes these mutants.
- FGF2 receptor means a cell surface protein to which FGF2 binds.
- FGF2 receptor FGFR1b, FGFR1c, FGFR2c, FGFR3c, and FGFR4 are known.
- the FGF2 receptor in the present invention may be a protein containing a natural amino acid sequence or a variant thereof.
- the “mutant” of the FGF2 receptor is one in which one to several amino acids are substituted, deleted, added, or the like, or a partial amino acid sequence of the known FGF2 receptor amino acid sequence.
- the present invention provides an aptamer that inhibits the binding of FGF2 to the FGF2 receptor.
- the aptamer of the present invention may have an inhibitory activity against FGF2 derived from any mammal.
- mammals include, for example, primates (eg, humans, monkeys), rodents (eg, mice, rats, guinea pigs), and pets, livestock and working animals (eg, dogs, cats, horses, cows). , Goats, sheep, pigs).
- the aptamer of the present invention may have a feature that it can inhibit the activity of FGF2, but cannot inhibit the activity of FGF1. In one embodiment, the aptamer of the present invention may be characterized by inhibiting binding between FGF2 and FGF2 receptor but not inhibiting binding between FGF1 and FGF1 receptor.
- FGF1 is an FGF family protein and is most similar to FGF2.
- N 1 and N 6 each independently represent any 0 to several bases, and N 2 , N 3 , N 4 and N 5 are independently any one base.
- the “base” means any one of adenine (A), guanine (G), cytosine (C), uracil (U), and thymine (T) constituting nucleic acid.
- the number of bases of N 1 is not particularly limited as long as the aptamer including the nucleotide sequence represented by formula (1) binds to FGF2, but, for example, 0 to about 10, 0 to 9, 0 to 8, 0 to It may be 7, 0-6, 0-5, 0-4, 0-3, 0-2, etc., preferably 0-2.
- the number of N 6 bases is not particularly limited as in N 1 , but for example, 0 to about 10, 0 to 9, 0 to 8, 0 to 7, 0 to 6, 0 to 5, It may be 0-4, 0-3, etc., preferably 0-10, 3-9, or 5-8.
- N 1 is G, GG, AG, C or a gap
- N 2 is A or U
- N 3 is G, C or A
- N 4 is G, C or U
- N 5 is G or U
- N 6 is UUCN 61 or AGUCN 62 (wherein N 61 and N 62 are each independently any 0 to several bases).
- N 1 is the "gap", that no N 1 is present in the formula (1), i.e. N 1 is meant to be a 0 bases.
- the number of bases of N 61 is not particularly limited, but may be, for example, 0 to about 10, 0 to 7, 0 to 6, 0 to 5, 0 to 4, etc., preferably 0 to 5 There may be 1-5, or 2-4.
- the number of bases of N 62 is not particularly limited, and may be, for example, 0 to about 10, 0 to 7, 0 to 5, 0 to 4, 0 to 3, etc., preferably 0 to 5 It can be 0, 4 or 0, or 3.
- N 1 is G, GG, AG or a gap
- N 2 is A or U
- N 3 is G or A
- N 4 is C or U
- N 5 is G or U
- N 6 is UUCN 61 or AGUCN 62 (wherein N 61 and N 62 are as defined above).
- the aptamer of the present invention has the following formula (2) or (3): GGGAAACUAGGGCGUUAACGUGACCAGGUGUUCN 61 (Formula 2) N 1 GGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCN 62 (Formula 3) (Wherein N 1 , N 61 and N 62 are as defined above)
- the aptamer of the present invention comprises the nucleotide sequence represented by any of SEQ ID NOs: 1-12.
- the nucleotide sequences represented by SEQ ID NOs: 1 to 12 are shown below (however, uracil may be thymine) (hereinafter, A, G, C and U are nucleotide bases of adenine, guanine and cytosine, respectively) And uracil): SEQ ID NO: 1: GGGAUACUAAGGGCAAUUAAUGUUACCAGUGUAGUUCUCGA, SEQ ID NO: 2: GGGAAACUAGGGCGUUAACGUGACCAGGUGUUCUCGA, SEQ ID NO: 3 GGGAUACUAAGGGCAAUUAAUGUUACCAGGUGUAGUCCC, SEQ ID NO: 4: GGAUACUAGGGGAUAUAUGUUACCAGGUAGUCC, SEQ ID NO: 5: GGGGAAUCAUGGGCAAUUAAUGUUACCAGGUGUAGUCCCC
- the aptamer of the present invention comprises the nucleotide sequence represented by SEQ ID NO: 1, 3, 4, 5, 6, 8, 10 or 11.
- the aptamer of the present invention comprises the nucleotide sequence represented by SEQ ID NO: 2 or 7.
- the aptamer of the present invention comprises the nucleotide sequence represented by SEQ ID NO: 1, 3, 4, 5 or 6.
- the aptamer of the present invention may comprise a nucleotide sequence in which one or several nucleotides are substituted, deleted, inserted or added in any of the nucleotide sequences described above as long as they still bind to FGF2.
- a nucleotide contained in the aptamer (I) the ribose 2 'position of each pyrimidine nucleotide is a fluorine atom; (Ii) an aptamer in which the ribose 2 ′ position of each purine nucleotide is a hydroxy group; (B) in the aptamer of (a), (I) The 2'-position fluorine atom of ribose of each pyrimidine nucleotide is each independently unsubstituted or substituted with an atom or group selected from the group consisting of a hydrogen atom, a hydroxy group and a methoxy group , (Ii) The 2'-position hydroxy group of ribose of each purine nucleotide is independently unsubstituted or substituted with an atom or group selected from the group consisting of a hydrogen atom, a methoxy group and a fluorine atom It may be an aptamer
- the number of nucleotides to be substituted, deleted, inserted or added is not particularly limited as long as it still binds to FGF2 after substitution, deletion, insertion or addition.
- 1 to about 10, preferably 1 to The number may be 6, more preferably 1 to 5, further preferably 1 to 4, further preferably 1 to 3, and most preferably 1 or 2.
- the site where the nucleotide is substituted, deleted, inserted or added is not particularly limited as long as it still binds to FGF2 after substitution, deletion, insertion or addition, but in the above formulas (1), (2) and (3) In the site specified for one kind of nucleotide, nucleotides are substituted, deleted, inserted or added at 1 to 3, preferably 1 or 2, more preferably 1 site.
- nucleotides for example, 1 to about 10, preferably 1 to 6, more preferably, are present at a site where multiple types of nucleotides can be taken. 1 to 5, more preferably 1 to 4) substitutions, deletions, insertions or additions may be allowed.
- the length of the aptamer of the present invention is not particularly limited, and can usually be about 10 to about 200 nucleotides, for example, about 20 nucleotides or more (eg, 25 nucleotides or more, 30 nucleotides or more, 31 nucleotides or more, 32 nucleotides or more). 33 nucleotides or more), preferably 25 nucleotides or more, more preferably 30 nucleotides or more, and even more preferably 33 nucleotides or more.
- nucleotides or less usually about 80 nucleotides or less, preferably about 70 nucleotides or less, more preferably about 60 nucleotides or less, more preferably about 50 nucleotides or less, further preferably about 45 nucleotides or less (eg, 44 nucleotides) Or less, 43 nucleotides or less, 42 nucleotides or less, 41 nucleotides or less, 40 nucleotides or less). If the total number of nucleotides is small, chemical synthesis and mass production are easier, and the merit in cost is also great. In addition, chemical modification is easy, the in vivo stability is high, and the toxicity is considered low.
- the length of the aptamer of the present invention can usually be about 10 to about 200 nucleotides, preferably 20 to 80 nucleotides, more preferably 25 to 60 nucleotides, and further preferably 25 to 50 nucleotides. Most preferably, it is 30 to 45 nucleotides.
- the aptamer of the present invention also includes a plurality of aptamers (aptamer (A)) including a nucleotide sequence represented by the above formula (1), one or several nucleotides in the nucleotide sequence represented by the above formula (1)
- a plurality of aptamers (aptamer (B)) containing nucleotide sequences in which is substituted, deleted, inserted or added, and one or a plurality of aptamers (A) and one or a plurality of aptamers (B) May be a connected product selected from the group consisting of These conjugates can also bind to FGF2.
- the connection can be made in a tandem connection.
- a linker may be used for the connection.
- Linkers include nucleotide chains (eg, 1 to about 20 nucleotides), non-nucleotide chains (eg, — (CH 2 ) n -linker, — (CH 2 CH 2 O) n -linker, hexaethylene glycol linker, TEG linker , A linker containing a peptide, a linker containing a —S—S— bond, a linker containing a —CONH— bond, and a linker containing a —OPO 3 — bond).
- the number of the plurality of connected objects is not particularly limited as long as it is 2 or more, but may be 2, 3, or 4, for example.
- Each nucleotide contained in the aptamer of the present invention is the same or different and is a nucleotide containing a hydroxy group at the 2 ′ position of ribose (eg, ribose of pyrimidine nucleotide, ribose of purine nucleotide) (ie, natural ribonucleotide).
- ribose eg, ribose of pyrimidine nucleotide, ribose of purine nucleotide
- it may be a nucleotide (which may be referred to as “modified nucleotide” in the present invention) in which the hydroxy group is substituted (modified) with any atom or group at the 2 ′ position of ribose.
- Examples of such an arbitrary atom or group include a hydrogen atom, a fluorine atom, —O-alkyl group (eg, —O—Me group), —O-acyl group (eg, —O—CHO group), amino And nucleotides substituted with groups (eg, —NH 2 groups).
- the aptamers of the present invention also have at least one (eg, 1, 2, 3 or 4) nucleotides at the 2 ′ position of ribose, a hydroxy group, or any atom or group described above, eg, a hydrogen atom, It may be a modified nucleotide containing at least two types (for example, 2, 3 or 4 types) of groups selected from the group consisting of a fluorine atom, a hydroxy group and an —O—Me group.
- all the pyrimidine nucleotides are nucleotides in which the 2′-position of ribose is a fluorine atom, or the fluorine atoms are the same or different and are not substituted. It may be a nucleotide substituted with an atom or group, preferably an atom or group selected from the group consisting of a hydrogen atom, a hydroxy group and a methoxy group.
- aptamers in which the ribose 2 ′ position of all pyrimidine nucleotides is fluorinated are obtained.
- the aptamer of the present invention in which the fluorine atom is substituted with other atoms or groups described above can be produced by the method described below.
- all the purine nucleotides are nucleotides in which the 2′-position of ribose is a hydroxy group, or the hydroxy groups are the same or different and unsubstituted, It may be a nucleotide substituted with an atom or group, preferably an atom or group selected from the group consisting of a hydrogen atom, a methoxy group and a fluorine atom.
- the aptamer of the present invention in which the hydroxy group is substituted with the other atom or group can be produced by the method described below.
- all pyrimidine nucleotides are selected from the group consisting of the above-described atoms or groups in which the fluorine atom at the 2′-position of ribose is a hydrogen atom, a hydroxy group, and an —O—Me group. It may be a nucleotide substituted with the same atom or group.
- all the purine nucleotides are selected from the group consisting of the above-described atoms or groups, for example, a hydrogen atom, a fluorine atom and an —O-Me group, in which the 2′-position hydroxy group of ribose It may be a nucleotide substituted with the same atom or group.
- each pyrimidine nucleotide contained in the aptamer of the present invention is a nucleotide containing a fluorine atom at the 2 ′ position of ribose, and each purine nucleotide has a hydroxy group at the 2 ′ position of ribose. Containing nucleotides.
- the 2'-position fluorine atom of ribose of each pyrimidine nucleotide may be independently substituted with an atom or group selected from the group consisting of a hydrogen atom, a hydroxy group and a methoxy group
- the hydroxyl group at the 2 ′ position of ribose of each of the purine nucleotides may be independently substituted with an atom or group selected from the group consisting of a hydrogen atom, a methoxy group and a fluorine atom.
- the mode of modification to the sugar group in the nucleotide will be described. Does not mean that DNA is excluded from the nucleotides constituting, and is appropriately read as a modification to DNA.
- the nucleotide constituting the aptamer is DNA
- the replacement of the hydroxyl group at the 2 ′ position of ribose with X is read as the replacement of the hydrogen atom at the 2 ′ position of deoxyribose with X.
- the aptamer of the present invention by replacing uracil with thymine, the binding ability to FGF2, the binding inhibitory activity between FGF2 and the FGF receptor, the stability of the aptamer, the drug delivery property, the stability in blood, etc. are enhanced. Is possible.
- one or several phosphodiester bonds in nucleotides are modified with an arbitrary substituent. Or it may be substituted.
- the phosphodiester bond may be substituted with a phosphorothioate bond, a phosphorodithioate bond, an alkylphosphonate bond, a phosphoramidate bond, or the like.
- a nucleotide is substituted with a phosphorothioate bond means that the phosphate group at the binding site between adjacent nucleotides is sulfurated, that is, the phosphodiester bond is modified to a phosphorothioate bond. It shows that.
- nucleic acid for the purpose of stabilizing the aptamer and improving its activity, 1 or several, for example, 1-2, 1-3, 1-4, 1-5 nucleotides are contained. It may be substituted with a crosslinked nucleic acid Bridged Nucleic Acid (BNA) or Locked Nucleic Acid (LNA).
- BNA Bridged Nucleic Acid
- LNA Locked Nucleic Acid
- the “crosslinked nucleic acid” refers to a nucleic acid having a structure that increases binding affinity to a complementary sequence and acquires nuclease resistance by constraining the degree of freedom of the nucleic acid by intramolecular crosslinking. Examples include, but are not limited to, ', 4'-BNA (LNA), 2'-O, 4'-C-ethylene-bridgedENNucleic Acid (ENA).
- the aptamer of the present invention is an aptamer that binds to FGF2, more preferably an aptamer that can bind to FGF2 and inhibit the binding between FGF2 and the FGF receptor. Whether or not the aptamer of the present invention inhibits the binding between FGF2 and the FGF receptor can be evaluated by, for example, a test using a surface plasmon resonance method such as Example 1.
- the aptamer of the present invention may be one in which the sugar residue (eg, ribose) of each nucleotide is modified in order to enhance the binding property to FGF2, stability, drug delivery property and the like.
- the site modified in the sugar residue include those in which the oxygen atom at the 2′-position, 3′-position and / or 4′-position of the sugar residue is replaced with another atom.
- modifications include fluorination, O-alkylation (eg, O-methylation, O-ethylation), O-allylation, S-alkylation (eg, S-methylation, S-ethylation). ), S-allylation, amination (eg, —NH 2 ).
- the aptamer of the present invention may be produced from the production method with a certain modification of the ribose 2′-position oxygen atom of the pyrimidine nucleotide, and a production method using DuraScribe TM T7 Transcription Kit (produced by Epicentre), which will be described later.
- the aptamer of the present invention can be an aptamer in which at least one nucleotide sugar residue is modified.
- modification of sugar residues can be carried out by a method known per se (for example, Sproat et al., (1991), Nucl. Acid. Res. 19, 733-738; Cotton et al., (1991)). ), Nucl.Acid.Res.
- the hydroxyl group at the ribose 2 ′ position is selected from the group consisting of a hydrogen atom, a hydroxyl group and a methoxy group based on an aptamer in which the ribose 2 ′ position hydroxyl group of all pyrimidine nucleotides is substituted with a fluoro group. Aptamers substituted with atoms or groups can be produced.
- the aptamer of the present invention may also be one in which a nucleobase (eg, purine or pyrimidine) is modified (eg, chemically substituted) in order to enhance binding properties to FGF2, stability, drug delivery properties, and the like.
- a nucleobase eg, purine or pyrimidine
- modifications include 5-position pyrimidine modification, 6- and / or 8-position purine modification, modification with exocyclic amines, substitution with 4-thiouridine, and substitution with 5-bromo or 5-iodo-uracil.
- the phosphate group contained in the aptamer of the present invention may be modified so as to be resistant to nuclease and hydrolysis.
- the P (O) O group may be P (O) S (thioate), P (S) S (dithioate), P (O) N (R) R ′ (amidate), P (O) R, P ( O) optionally substituted by OR, CO or CH 2 (formacetal) or 3′-amine (—NH—CH 2 —CH 2 —), wherein each R or R ′ is independently H Or substituted or unsubstituted alkyl (eg, methyl, ethyl)].
- the linking group include —O—, —N—, and —S—, which can be bonded to adjacent nucleotides through these linking groups. Modifications may also include 3 ′ and 5 ′ modifications such as capping.
- Modifications are further made of polyethylene glycol, amino acids, peptides, inverted dT, nucleic acids, nucleosides, Myristoy, Lithocolic-oleyl, Docosanyl, Lauroyl, Stearoyl, Palmitoyl, Oleoyl, Linoleyl, other lipids, vitamins, steroids, cholesterol, steroids It can be performed by adding a fluorescent substance, an anticancer agent, a toxin, an enzyme, a radioactive substance, biotin or the like to the terminal. See US Pat. Nos. 5,660,985 and 5,756,703 for such modifications.
- the molecular weight of PEG is not particularly limited, but is preferably 1,000 to 100,000, more preferably 30,000 to 90,000.
- PEG may be linear or may be branched into two or more chains (multi-arm PEG).
- Such PEG is not particularly limited, and those skilled in the art can appropriately select and use commercially available or known PEG (for example, http://www.peg-drug.com/peg_product/branched.html).
- PEG to be applied to the aptamer of the present invention specifically, a bibranched GS type PEG having a molecular weight of 40000 (SUNBRIGHT GL2-400GS manufactured by NOF Corporation), a bibranched TS type PEG having a molecular weight of 40000 (SUNBRIGHT GL2-400TS made by NOF), 4-branched TS type PEG having a molecular weight of 40000 (SUNBRIGHT GL4-400TS made by NOF), bibranched TS type PEG having a molecular weight of 80000 (SUNBRIGHT GL2-800TS made by NOF), or a molecular weight of 80000 4 branches S-type PEG (manufactured by SUNBRIGHT GL4-800TS Date oil), and the like.
- PEG may be directly added to the terminal, but a linker having a group capable of binding to PEG is added to the terminal, through which PEG is added to the aptamer of the present invention. It is more preferable to add.
- the linker between PEG and the aptamer of the present invention is not particularly limited, and the number of carbon chains, functional groups, and the like can be appropriately selected according to the binding site, the type of PEG, and the like.
- Examples of such a linker include a linker having an amino group.
- SAFC ssH Linker
- DMS O
- MT-AMINO-MODIFIER GLENRESEARCH
- TFA Amino C-6 lcaa CPG (ChemGenes) is exemplified.
- this linker for example, an N-hydroxysuccinimide active group is added to PEG, and this is reacted with an amino group on the linker side, thereby binding the aptamer of the present invention and PEG via the linker. can do.
- PEG and a linker a commercially available thing can be used preferably.
- those skilled in the art can appropriately set reaction conditions relating to the binding of PEG, a linker, and the aptamer of the present invention.
- the aptamer of the present invention can be chemically synthesized by the disclosure in the present specification and a method known per se in the technical field.
- Aptamers bind to a target substance by various binding modes such as ionic bonds using the negative charge of the phosphate group, hydrophobic bonds and hydrogen bonds using ribose, hydrogen bonds using nucleobases and stacking bonds.
- the ionic bond utilizing the negative charge of the phosphate group that exists in the number of constituent nucleotides is strong and binds to the positive charge of lysine or arginine present on the surface of the protein. For this reason, nucleobases that are not involved in direct binding to the target substance can be substituted.
- the stem structure portion is already base-paired and faces the inside of the double helix structure, so that the nucleobase is difficult to bind directly to the target substance. Therefore, the activity of the aptamer is often not reduced even if the base pair is replaced with another base pair. Even in a structure that does not form a base pair, such as a loop structure, base substitution is possible when the nucleobase is not involved in direct binding to the target molecule.
- the modification of the 2 'position of ribose in rare cases, the functional group at the 2' position of ribose may directly interact with the target molecule, but in many cases it is irrelevant and can be replaced with other modified molecules. Is possible. Thus, aptamers often retain their activity unless the functional group involved in direct binding to the target molecule is replaced or deleted. It is also important that the overall three-dimensional structure does not change significantly.
- aptamers use the SELEX method and its improved methods (for example, Ellington et al., (1990), Nature, 346, 818-822; Tuerk et al., (1990), Science, 249, 505-510). Can be produced.
- SELEX method by increasing the number of rounds or using a competitive substance, aptamers having a stronger binding force with respect to the target substance are concentrated and selected. Therefore, by adjusting the number of rounds of SELEX and / or changing the competition state, aptamers with different binding strengths, aptamers with different binding modes, binding strengths and binding modes are the same, but base sequences are different. Aptamers may be obtained.
- the SELEX method includes an amplification process by PCR. By introducing a mutation by using manganese ions in the process, it becomes possible to perform SELEX with more diversity.
- An aptamer obtained by SELEX is a nucleic acid having a high affinity for a target substance, which does not mean binding to the active site of the target substance. Therefore, the aptamer obtained by SELEX does not necessarily affect the function of the target substance.
- FGF2 is a basic protein, and it is thought that nucleic acids are likely to bind nonspecifically. Aptamers that do not bind to the active site do not affect the activity of the target substance. In fact, the RNA used as a control did not inhibit the binding of FGF2 and FGF2 receptor.
- the active aptamer thus selected can be further improved in performance by performing the optimized SELEX.
- the optimized SELEX a template in which a part of an aptamer having a predetermined sequence is made into a random sequence or a template in which about 10 to 30% of a random sequence is doped is prepared, and SELEX is performed again.
- the aptamer obtained by SELEX has a length of about 80 nucleotides, and it is difficult to make it as a medicine as it is. Therefore, trial and error are repeated, and the length is reduced to a length that allows easy chemical synthesis (for example, chemical synthesis can be performed at about 60 nucleotides or less, more preferably about 50 nucleotides or less, and even more preferably 45 nucleotides or less). It is preferable.
- the aptamers obtained by SELEX vary in ease of subsequent minimization work depending on the primer design. If the primer is not designed well, even if active aptamers can be selected by SELEX, subsequent development becomes impossible.
- Aptamers are easy to modify because they can be chemically synthesized. Aptamers use the MFOLD program to predict secondary structures, predict three-dimensional structures by X-ray analysis or NMR analysis, which nucleotides can be replaced or deleted, and where new nucleotides can be found. It can be predicted to some extent whether or not can be inserted. Aptamers of the predicted new sequence can be easily chemically synthesized, and whether or not the aptamer retains activity can be confirmed by an existing assay system.
- the activity often changes even if a new sequence is added to both ends of the sequence. do not do.
- the length of the new sequence is not particularly limited.
- the modification can be designed or altered to a high degree in the same manner as the sequence.
- aptamers can be highly designed or modified.
- the present invention also includes a predetermined sequence (eg, a sequence corresponding to a portion selected from a stem portion, an internal loop portion, a hairpin loop portion, and a single-stranded portion: hereinafter, abbreviated as a fixed sequence if necessary).
- a predetermined sequence eg, a sequence corresponding to a portion selected from a stem portion, an internal loop portion, a hairpin loop portion, and a single-stranded portion: hereinafter, abbreviated as a fixed sequence if necessary.
- a method for producing an aptamer in which the aptamer can be highly designed or modified.
- the method for producing such aptamer is as follows:
- (N) a represents a nucleotide chain consisting of a N
- (N) b represents a nucleotide chain consisting of b N
- N is the same or different, respectively, A, G
- It is a nucleotide selected from the group consisting of C, U and T (preferably A, G, C and U).
- a and b are the same or different and may be any number, for example, 1 to about 100, preferably 1 to about 50, more preferably 1 to about 30, even more preferably 1 to about 20 or 1
- a single nucleic acid molecule or a plurality of nucleic acid molecules eg, a library of nucleic acid molecules having different numbers of a, b, etc.
- a primer sequence (i ) And (ii) respectively, to produce an aptamer containing a fixed sequence using a primer pair corresponding to each.
- the aptamer of the present invention is preferably one of the following (a ′), (b ′) or (c ′), which binds to FGF2 and inhibits binding between FGF2 and the FGF receptor: (a ′ ) Including a nucleotide sequence represented by any one of SEQ ID NOS: 1 to 7 (or SEQ ID NO: 2 or 7, or any of SEQ ID NOS: 1 and 3 to 6) (however, uracil may be thymine)
- the present invention also provides a complex comprising the aptamer of the present invention and a functional substance bound thereto.
- the bond between the aptamer and the functional substance in the complex of the present invention can be a covalent bond or a non-covalent bond.
- the complex of the present invention may be a conjugate of the aptamer of the present invention and one or more (eg, 2 or 3) of the same or different functional substances.
- the functional substance is not particularly limited as long as it newly adds some function to the aptamer of the present invention or can change (eg, improve) some property that can be retained by the aptamer of the present invention.
- Examples of the functional substance include proteins, peptides, amino acids, lipids, carbohydrates, monosaccharides, polynucleotides, and nucleotides.
- Examples of functional substances include, for example, affinity substances (eg, biotin, streptavidin, polynucleotides having affinity for target complementary sequences, antibodies, glutathione sepharose, histidine), labeling substances (eg, fluorescent substances, Luminescent substances, radioisotopes), enzymes (eg, horseradish peroxidase, alkaline phosphatase), drug delivery vehicles (eg, liposomes, microspheres, peptides, polyethylene glycols), drugs (eg, calicheamicin and duocarmycin) Used in missile therapy, nitrogen mustard analogs such as cyclophosphamide, melphalan, ifosfamide or trophosphamide, ethyleneimines such as thiotepa, nitrosourea such as carmustine, temozo
- ⁇ унк ⁇ онент ⁇ may eventually be removed. Furthermore, it may be a peptide that can be recognized and cleaved by an enzyme such as thrombin, matrix metalloprotease (MMP), Factor X, or a polynucleotide that can be cleaved by a nuclease or a restriction enzyme.
- an enzyme such as thrombin, matrix metalloprotease (MMP), Factor X, or a polynucleotide that can be cleaved by a nuclease or a restriction enzyme.
- the aptamer or complex of the present invention can be used as, for example, a medicine or a diagnostic agent, a test agent, or a reagent.
- diseases associated with angiogenesis such as age-related macular degeneration, osteoporosis, rheumatoid arthritis, osteoarthritis, bone and cartilage diseases such as fractures, drugs for the treatment or prevention of pain, or diagnostic agents, test agents, Useful as a reagent.
- the medicament of the present invention may be formulated with a pharmaceutically acceptable carrier.
- the pharmaceutically acceptable carrier include excipients such as sucrose, starch, mannitol, sorbit, lactose, glucose, cellulose, talc, calcium phosphate, calcium carbonate, cellulose, methylcellulose, hydroxypropylcellulose, polypropylpyrrolidone , Gelatin, gum arabic, polyethylene glycol, sucrose, starch and other binders, starch, carboxymethylcellulose, hydroxypropyl starch, sodium-glycol starch, sodium bicarbonate, calcium phosphate, calcium citrate and other disintegrants, magnesium stearate , Aerosil, Talc, Lubricant such as sodium lauryl sulfate, Citric acid, Menthol, Glycyllysine / Ammonium salt, Glycine, Orange powder, etc.
- excipients such as sucrose, starch, mannitol, sorbit, lactose, glucose,
- Preservatives such as sodium, sodium bisulfite, methylparaben, propylparaben, stabilizers such as citric acid, sodium citrate, acetic acid, suspensions such as methylcellulose, polyvinylpyrrolidone, aluminum stearate, dispersants such as surfactants, Examples include, but are not limited to, water, physiological saline, diluents such as orange juice, base waxes such as cacao butter, polyethylene glycol, and white kerosene.
- Preparations suitable for oral administration include a solution in which an effective amount of a ligand is dissolved in a diluent such as water, physiological saline, orange juice, a capsule containing an effective amount of the ligand as a solid or a granule, a sachet or Examples thereof include tablets, suspensions in which an effective amount of an active ingredient is suspended in a suitable dispersion medium, and emulsions in which a solution in which an effective amount of an active ingredient is dissolved is dispersed in an appropriate dispersion medium and emulsified.
- a diluent such as water, physiological saline, orange juice
- a capsule containing an effective amount of the ligand as a solid or a granule a sachet or Examples thereof include tablets, suspensions in which an effective amount of an active ingredient is suspended in a suitable dispersion medium, and emulsions in which a solution in which an effective amount of an active ingredient is dissolved is dispersed
- the medicament of the present invention can be coated by a method known per se for the purpose of taste masking, enteric solubility or sustainability, if necessary.
- the coating agent used for coating include hydroxypropylmethylcellulose, ethylcellulose, hydroxymethylcellulose, hydroxypropylcellulose, polyoxyethylene glycol, Tween 80, Pluronic F68, cellulose acetate phthalate, hydroxypropylmethylcellulose phthalate, hydroxymethylcellulose acetate succinate, Eudragit (Rohm, Germany, methacrylic acid / acrylic acid copolymer) and pigments (eg, Bengala, titanium dioxide) are used.
- the medicine may be either an immediate release preparation or a sustained release preparation.
- the base material of the sustained release preparation include liposomes, atelocollagen, gelatin, hydroxyapatite, and PLGA.
- Suitable formulations for parenteral administration are aqueous and non-aqueous isotonic.
- parenteral administration eg, intravenous, subcutaneous, intramuscular, topical, intraperitoneal, nasal, pulmonary, etc.
- aqueous and non-aqueous isotonic are aqueous and non-aqueous isotonic.
- sterile injection solutions which may contain antioxidants, buffers, antibacterial agents, isotonic agents and the like.
- aqueous and non-aqueous sterile suspensions can be mentioned, which may contain suspending agents, solubilizers, thickeners, stabilizers, preservatives and the like.
- the preparation can be enclosed in a container in unit doses or multiple doses like ampoules and vials.
- the active ingredient and a pharmaceutically acceptable carrier can be lyophilized and stored in a state that may be dissolved or suspended in a suitable sterile solvent immediately before use.
- inhalants and ointments are also possible.
- the active ingredient in a lyophilized state is refined and administered by inhalation using an appropriate inhalation device.
- a conventionally used surfactant, oil, seasoning, cyclodextrin or a derivative thereof can be appropriately blended as necessary.
- surfactant examples include oleic acid, lecithin, diethylene glycol dioleate, tetrahydrofurfuryl oleate, ethyl oleate, isopropyl myristate, glyceryl trioleate, glyceryl monolaurate, glyceryl monooleate, glyceryl monostearate.
- Span, Tween, Epilon, Brij, Genapol, and Synperonic are trademarks.
- the oil include corn oil, olive oil, cottonseed oil and sunflower oil.
- an appropriate pharmaceutically acceptable base yellow petrolatum, white petrolatum, paraffin, plastibase, silicone, white ointment, beeswax, pig oil, vegetable oil, hydrophilic ointment, hydrophilic petrolatum, purified lanolin, hydrolyzed lanolin , Water-absorbing ointment, hydrophilic plastibase, macrogol ointment, etc.
- Inhalants can be manufactured according to conventional methods. That is, the aptamer or complex of the present invention can be produced by making it powder or liquid, blending it in an inhalation propellant and / or carrier, and filling it in an appropriate inhalation container.
- the aptamer or complex of the present invention is a powder, a normal mechanical powder inhaler can be used, and when it is a liquid, an inhaler such as a nebulizer can be used.
- the propellant conventionally known ones can be widely used.
- the medicament of the present invention can be administered not only directly to the lesion site but also by other methods described above.
- the aptamer of the present invention is a single-stranded nucleic acid, it can be detoxified by administration of a nucleotide containing a complementary sequence, and may be a safer pharmaceutical than a neutralizing antibody whose kinetic control is difficult after administration. high. This can be said to be extremely advantageous in view of the problem of infectious diseases caused by the long residence time of the antibody in the body, which can occur in antibody drug therapy.
- the medicament of the present invention when used as a medicament for the prevention or treatment of the above-mentioned diseases, considering the seriousness of the disease and the risk of side effects, it is safer to use aptamers that easily control pharmacokinetics. It is clear that a medicine can be obtained.
- the dose of the medicament of the present invention varies depending on the type / activity of the active ingredient, the severity of the disease, the animal species to be administered, the drug acceptability of the administration target, body weight, age, etc.
- the amount of active ingredient may be about 0.0001 to about 100 mg / kg, such as about 0.0001 to about 10 mg / kg, preferably about 0.005 to about 1 mg / kg.
- the aptamer or complex of the present invention can also be used as a drug delivery agent, an in vivo imaging probe, an FGF2 blood concentration measurement probe, a tissue staining probe, an ELISA probe, and a ligand for separation and purification of FGF2.
- the present invention also provides a solid phase carrier on which the aptamer or complex of the present invention is immobilized.
- the solid phase carrier include a substrate, a resin, a plate (eg, multiwell plate), a filter, a cartridge, a column, and a porous material.
- the substrate may be one used for DNA chips, protein chips, etc., for example, nickel-PTFE (polytetrafluoroethylene) substrate, glass substrate, apatite substrate, silicone substrate, alumina substrate, etc. And the like coated with a polymer or the like.
- the resin examples include agarose particles, silica particles, copolymers of acrylamide and N, N′-methylenebisacrylamide, polystyrene-crosslinked divinylbenzene particles, particles obtained by crosslinking dextran with epichlorohydrin, cellulose fibers, and allyldextran.
- examples include N, N'-methylenebisacrylamide cross-linked polymers, monodisperse synthetic polymers, monodisperse hydrophilic polymers, sepharose, and Toyopearl.
- resins in which various functional groups are bonded to these resins. .
- the solid phase carrier of the present invention can be useful for, for example, purification of FGF2, and detection and quantification of FGF2.
- the aptamer or complex of the present invention can be immobilized on a solid support by a method known per se.
- an affinity substance eg, those described above
- a predetermined functional group is introduced into the aptamer or complex of the present invention, and then immobilized on a solid phase carrier using the affinity substance or the predetermined functional group.
- a method is mentioned.
- the present invention also provides a method for immobilizing the aptamer or complex of the present invention to a solid support, and the solid support thus obtained.
- the predetermined functional group may be a functional group that can be subjected to a coupling reaction, and examples thereof include an amino group, a thiol group, a hydroxy group, and a carboxyl group.
- the present invention also provides an aptamer having such a functional group introduced therein.
- the present invention also provides a method for purification and concentration of FGF2.
- the purification method of the present invention can separate FGF2 from other FGF family proteins.
- the purification and concentration method of the present invention can include adsorbing FGF2 on the solid phase carrier of the present invention and eluting the adsorbed FGF2 with an eluent.
- Adsorption of FGF2 to the solid phase carrier of the present invention can be carried out by a method known per se. For example, a sample containing FGF2 (eg, bacterial or cell culture or culture supernatant, blood) is introduced into the solid phase carrier of the present invention or its contents. The elution of FGF2 can be performed using an eluent such as a neutral solution.
- the neutral eluate is not particularly limited, and may be, for example, a pH of about 6 to about 9, preferably about 6.5 to about 8.5, more preferably about 7 to about 8.
- Neutral solutions can also include, for example, potassium salts (eg, KCl), magnesium salts (eg, MgCl 2 ), surfactants (eg, Tween 20, Triton, NP40), glycerin.
- the purification and concentration method of the present invention may further comprise washing the solid phase carrier with a washing solution after FGF2 adsorption.
- the cleaning liquid examples include urea, a chelating agent (eg, EDTA), Tris, acid, alkali, transfer RNA, DNA, a surfactant such as Tween 20, and a salt containing a salt such as NaCl.
- the purification and concentration method of the present invention may further include heat-treating the solid phase carrier. By this process, the solid phase carrier can be regenerated and sterilized.
- the aptamer or complex of the present invention can be used as a detection probe, particularly a detection probe for FGF2.
- the labeling method of the aptamer is not particularly limited, and a method known per se can be applied. Examples of such a method include labeling with a radioisotope, labeling with a fluorescent dye or a fluorescent protein, and the like.
- the present invention also provides a method for detecting and quantifying FGF2.
- the present invention can detect and quantify FGF2 separately from other FGF family proteins.
- the detection and quantification method of the present invention can include measuring FGF2 utilizing the aptamer of the present invention (eg, by using the complex of the present invention and a solid phase carrier).
- the method for detecting and quantifying FGF2 can be performed by the same method as the immunological method except that the aptamer of the present invention is used instead of the antibody.
- an enzyme immunoassay eg, direct competition ELISA, indirect competition ELISA, sandwich ELISA
- radioimmunoassay RIA
- fluorescence immunoassay FIA
- Western blotting immunohistochemical staining, cell sorting, and similar methods
- a method can be useful, for example, for measuring FGF2 levels in biological or biological samples, diagnosing FGF2-related diseases.
- Example 1 Preparation of RNA aptamer specifically binding to FGF2
- a library with about 20 mer primers attached to both ends of a random sequence of about 30 mer to 40 mer was used.
- the total length of the obtained aptamer was about 80 to 100 mer, and it was necessary to shorten the chain thereafter.
- shortening the chain is not always easy, and the activity often decreases greatly. Therefore, with reference to the Taylored-SELEX method developed by NOXXON (Veter et al. Nucleic Acids Res. 31, 2003, el30; Jarosch et al. Nucleic Acids Res.
- DNA template 5′-TCGAG-30N-TCCCTATAGTGAGTCGTATTAGCAGCTCCCACAGGCTT-3 ′ (SEQ ID NO: 13)
- Forward ligate 5′-UAAUACGACUUCACUAUA-3 ′
- Forward primer 5′-AAGCCTGTGGAGCTGCTAATACGACTCACTATAGGGA-3 ′
- Forward bridge 5′-TCCCTATAGTGAGTCGTATTA-NH2-3 ′
- Reverse bridge 5′-TCTTGTTCAGCTTAGTTCTCTCGAG-3 ′ (SEQ ID NO: 17)
- Reverse ligate 5′-p-GAGAACTAAGCTGAACAAGA-NH2-3 ′ (SEQ ID NO: 18)
- FGF2 Human FGF2 (manufactured by Peprotech) was used as a target substance. FGF2 was immobilized on an agarose resin (NHS-activated Sepharose, manufactured by GE Healthcare) by amino coupling. Amino coupling was performed according to the specifications of GE Healthcare. The amount of immobilization was confirmed by examining the FGF2 solution before immobilization and the supernatant immediately after immobilization by SDS-PAGE. As a result of SDS-PAGE, no FGF2 band was detected in the supernatant, confirming that almost all of the FGF2 used was coupled. About 290 pmol of FGF2 was immobilized on about 5 ⁇ L of resin.
- RNA (30N-RNA) used in the first round was transcribed using a DNA template obtained by chemical synthesis into double strands using Forward primer, and using DuraScribe TM T7 Transcribation Kit (manufactured by Epicentre). I got it.
- the RNA obtained by this method is one in which the 2'-position of the ribose of the pyrimidine nucleotide is fluorinated. After two rounds, the DNA was double-stranded, and then the 3 'primer sequence was cleaved with a restriction enzyme before transcription.
- RNA bound to FGF2 was recovered at 95 ° C. for 10 minutes after adding the eluate.
- 7 M Urea, 3 mM EDTA and 0.1 M Tris were mixed and adjusted to pH 6.6.
- the PCR product was cloned into pGEM-T Easy vector (Promega) and transformed into E. coli strain DH5 ⁇ (Toyobo). After extracting the plasmid from the single colony, the base sequence was examined with a DNA sequencer (3130xl Genetic Analyzer, manufactured by ABI).
- the binding activity of the nucleic acids represented by SEQ ID NOs: 1 and 2 to FGF2 was evaluated by the surface plasmon resonance method.
- the nucleotide sequences represented by SEQ ID NOs: 1 and 2 are shown as aptamer IDs 1 and 2 together with the modification at the 2 ′ position of ribose.
- the brackets in each nucleotide indicate the modification at the 2 ′ position of ribose, and F indicates a fluorine atom.
- C (F) represents cytidine in which the 2 ′ position of ribose is substituted with a fluorine atom
- U (F) represents uridine in which the 2 ′ position of ribose is substituted with a fluorine atom.
- the beginning of each sequence is the 5 ′ end and the tail is the 3 ′ end.
- Aptamer ID 1 GGGAU (F) AC (F) U (F) AGGGGC (F) AU (F) U (F) AAU (F) GU (F) U (F) AC (F) C (F) AGU (F) GU ( F) AGU (F) C (F) U (F) C (F) GA
- Aptamer ID 2 GGGAAAC (F) U (F) AGGGGC (F) GU (F) U (F) AAC (F) GUC (F) GAC (F) C (F) AGU (F) GU (F) U (F) U (F) C (F) GA
- BIAcore2000 manufactured by BIAcore was used for measurement, and CM4 that reacts with amino groups was used as a sensor chip.
- Human FGF2 was dissolved in an immobilization solution (10 mM sodium acetate, pH 6) to a concentration of 25-40 ⁇ g / mL. Ethyl-3-carbohydrate hydrochloride and N-hydroxysuccinimide were used for the reaction of the amino group on the protein side and the carboxyl group on the chip side. After the reaction, blocking with ethanolamine-HCl was performed. The amount of FGF2 immobilized was 2500 to 4000 RU.
- the aptamer for the analyte was prepared at 0.15 ⁇ M to 0.5 ⁇ M. Solution A was used as the running buffer. 2M NaCl was used as a regeneration solution. FGF2 was immobilized on the flow cell F C 2 and the final sensorgram was obtained by subtracting the result of F C 1.
- FIG. 1 shows a sensorgram showing the state of binding between the aptamer represented by the aptamers ID1 and 2 and human FGF2.
- BIAcore2000 manufactured by BIAcore was used for the measurement.
- Protein A manufactured by PIERCE
- Human FGFR1 ⁇ (IIIc), R2 ⁇ (IIIc), R3 (IIIc), and R4 (R & D Systems, Inc.) fused with the Fc portion of IgG were each immobilized at about 1,000 RU.
- FIG. 2 shows a sensorgram showing that the aptamer represented by the aptamer ID1 inhibits the binding between FGF2 and FGFR1 ⁇ (IIIc), 2 ⁇ (IIIc), 3 (IIIc), and 4.
- the inhibition rates for each of the four types of receptors were determined.
- the maximum binding amount of FGF2 and heparin mixture was 0, and the binding amount when only the injection buffer was used was 100.
- the binding amount means the RU value at the peak top of the sensorgram.
- the aptamers represented by aptamer IDs 1 and 2 were as high as 50% or more for both receptors.
- the inhibition rate of other aptamers was 50% or less. Table 1 shows the results.
- Example 2 Shortening of the aptamer represented by SEQ ID NOs: 1 and 2
- the aptamer represented by SEQ ID NOs: 1 and 2 was shortened.
- the secondary structure of RNA was predicted using the MFOLD program (Zuker, Nucleic Acids Res. 31, 3406-3415, 2003), and shortened with reference to the structure.
- the shortened form was obtained by preparing DNA of the target sequence by chemical synthesis, and transcribing it using the DuraScript T7 Transcription Kit.
- the nucleotide sequences (SEQ ID NOs: 3 and 7) of the actually shortened products are shown below as aptamer IDs 3 and 7 together with the modification at the 2 ′ position of ribose.
- Aptamer ID3 A 36-nucleotide aptamer GGGAU (F) AC (F) U (F) AGGGGC (F) AU (F) U (F) AAU (F) GU ( F) U (F) AC (F) C (F) AGU (F) GU (F) AGU (F) C (F) C (F) C (F) Aptamer ID7: An aptamer variant represented by SEQ ID NO: 2 and having an aptamer length of 35 nucleotides GGGAAAC (F) U (F) AGGGGC (F) GU (F) U (F) AAC (F) GU (F) GAC ( F) C (F) AGU (F) GU (F) U (F) U (F) C (F) C (F) C (F) C (F) C (F) C (F) Aptamer ID7: An aptamer variant represented by SEQ ID NO: 2 and having an aptamer length of 35 nucleo
- Example 3 Specificity of the aptamer represented by aptamer ID3 Whether the FGF2 aptamer represented by aptamer ID3 binds to FGF1 of the same FGF family, or several growth factors EGF, ⁇ -NGF, VEGF Examined.
- BIAcore2000 manufactured by BIAcore was used for the measurement.
- the sensor chip used was an SA chip on which streptavidin was immobilized.
- About 500 RU was bound to the aptamer represented by aptamer ID3 to which biotin was added at the 5 terminal. Aptamers with biotin were produced by chemical synthesis.
- As the ligand protein FGF1, EGF, ⁇ -NGF, and VEGF manufactured by R & D were used.
- the running buffer used was a solution A used in Example 1 with sodium chloride added to a final concentration of 0.3M.
- aptamer represented by aptamer ID3 binds to FGF2 but does not bind to other proteins.
- the sensorgram is shown in FIG. From the above, it was found that the aptamer represented by aptamer ID3 specifically binds to FGF2.
- Example 4 Modification and modification of a shortened aptamer
- aptamer ID3 (1) -3 (40 ) In order to enhance the binding property to FGF2, stability, drug delivery, etc., aptamer ID3 (1) -3 (40 ), Aptamer IDs 4 and 4 (1) -4 (4), aptamer ID5 and aptamer ID6, nucleic acids were chemically synthesized.
- the aptamer represented by aptamer ID4 was obtained by cutting one of G (M) at the 5 ′ end and C (M) at the 3 ′ end of the aptamer represented by aptamer ID3 (19).
- the aptamer represented by aptamer ID5 was obtained by adding one G (M) at the 5 ′ end and one C (M) at the 3 ′ end of the aptamer represented by aptamer ID3 (19).
- the aptamer represented by aptamer ID6 was obtained by adding only A (M) to the 5 ′ end of the aptamer represented by aptamer ID3 (19).
- These nucleic acids were prepared by chemical synthesis. Whether the prepared aptamer inhibited the binding between FGF2 and the FGF receptor was examined in the same manner as in Example 1. Here, the concentrations of aptamer, FGF2, and heparin were all 0.1 ⁇ M. As a result of the experiment, it was found that all measured aptamers strongly inhibit the binding of FGF2 and FGFR1 ⁇ (IIIc) receptor. Table 3 shows the results.
- RNA Ribonucleic acid
- DNA Ribonucleic acid
- idT indicates inverted dT.
- the parentheses in each nucleotide indicate modification at the 2 ′ position
- F represents a fluorine atom
- M represents an O-methyl group.
- s represents a phosphorothioate bond.
- C6 represents — (CH 2 ) 6 -linker
- ssH represents ssH Linker (—CH 2 —CH 2 —O—CO—NH— (CH 2 ) 6 —).
- PEG40TS2 is a bibranched TS type polyethylene glycol having a molecular weight of 40000 (SUNBRIGHT GL2-400TS manufactured by NOF Corporation)
- PEG80TS4 is a 4-branched TS type having a molecular weight of 80000 (SUNBRIGHT GL4-800TS manufactured by NOF Corporation)
- Y-NHS-40K is a molecular weight of 40000.
- ME-100TS is a TS type with a molecular weight of 10,000 (SUNBRIGHT ME-100TS made by NOF Corporation), and PTE-100CS is a four-branch type with a molecular weight of 10,000 (SUNBRIGHT PTE-100CS manufactured by NOF Corporation).
- nucleotide sequence of aptamer ID3 (1)-(40) that does not contain a linker moiety or a modification moiety is represented by SEQ ID NO: 3, and similarly, aptamer ID4 and 4 (1)-(4), aptamer ID5 and The nucleotide sequences of 6 are represented by SEQ ID NOs: 4-6, respectively.
- Example 5 Evaluation of FGF2-dependent cell growth inhibitory activity of aptamers using human umbilical vein vascular endothelial cells
- Human umbilical vein vascular endothelial cells (HUVEC) were seeded at 5 ⁇ 10 3 cells per well in a 96-well flat bottom plate, and 2% bovine. The cells were cultured overnight in EGM-2 Bullet Kit (manufactured by CC-3162 Lonza) containing endothelial serum and growth factors. Thereafter, the medium was discarded, washed twice with PBS buffer, and aptamer (5, 2.5, 1, 0.5 nM) represented by aptamer ID3 (21) dissolved in a medium dedicated to endothelial cells containing 2% fetal bovine serum.
- EGM-2 Bullet Kit manufactured by CC-3162 Lonza
- aptamer represented by aptamer ID3 (21) inhibits angiogenesis.
- RNA As a negative control RNA, a Macugen (registered trademark) sequence without PEG added, which was C6 modified at the 5 'end and idT modified at the 3' end was used.
- the aptamer indicated by the aptamer ID also inhibits angiogenesis.
- Example 6 Evaluation of FGF2-dependent cell growth inhibitory activity 2 of aptamers using human umbilical vein vascular endothelial cells 2
- the inhibitory activity of aptamers represented by aptamer IDs 8 to 12 was measured.
- the results are shown in Table 6.
- the nucleotide sequences of aptamer IDs 8 to 12 are represented by SEQ ID NOs: 8 to 12, respectively.
- Example 7 Angiogenic mouse model test 0.76% (final concentration) sodium citrate-containing Matrigel (BD Matrigel TM ) containing human FGF-2 (manufactured by R & D) was added to 8-week-old C57BL / 6J mice ( ⁇ ) Under the anesthesia. Seven days later, Matrigel was removed, and the degree of neovascularization was evaluated by the amount of hemoglobin in Matrigel. The amount of hemoglobin was quantified by the cyanmethemoglobin method using Drabkin Reagent Kit. The aptamer was dissolved in a phosphate buffer containing 1 mM magnesium chloride and administered intraperitoneally once a day immediately after the subcutaneous administration of Matrigel.
- the administration groups are shown in Table 7, and the results are shown in Table 8. Significant angiogenesis inhibition was observed in the aptamer 1 mg / kg group. From the above, it was confirmed that the aptamer of the present invention exhibits strong angiogenesis inhibitory activity even in model animals.
- the aptamer or complex of the present invention may be useful as a medicament for a disease associated with angiogenesis, a disease such as a bone / cartilage disease or pain, or a diagnostic agent or reagent.
- the aptamers or complexes of the present invention may also be useful for FGF2 purification and enrichment, FGF2 labeling, and FGF2 detection and quantification.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Epidemiology (AREA)
- Physical Education & Sports Medicine (AREA)
- Rheumatology (AREA)
- Ophthalmology & Optometry (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Pain & Pain Management (AREA)
- Cardiology (AREA)
- Heart & Thoracic Surgery (AREA)
- Transplantation (AREA)
- Pulmonology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
Abstract
Description
FGF2は間葉系細胞を刺激してPDGFやPDGFR、VEGF、HGF等の炎症性サイトカインや増殖因子の発現を亢進させると共に、血管新生を促進し、骨破壊を促す。すなわちFGF2は、慢性関節リウマチにおける重要な病態に関与する中心的分子であることがわかっている(非特許文献1参照)。
FGF2の機能を制御する事ができれば、破骨細胞の活性化を介した関節破壊の治療薬としての効果も十分期待できると考えられ、実際に抗FGF2中和抗体をAIAモデルラットの関節に直接投与して症状が緩和することが知られている。しかしながらその発症の抑制効果はわずかであり、特に発症後の投与における治癒効果は確認されていない(非特許文献5)。
[1]下式(1)で表わされるヌクレオチド配列(但し、ウラシルはチミンであってもよい)を含むアプタマーであって、以下(a)又は(b)のいずれかである、FGF2に結合するアプタマー:
N1GGAN2ACUAGGGCN3UUAAN4GUN5ACCAGUGUN6(式1)
N1及びN6は、それぞれ独立して任意の0から数個の塩基、
N2、N3、N4及びN5は、独立して任意の一個の塩基を表す、
(a)該アプタマーに含まれるヌクレオチドにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位がフッ素原子であり、
(ii)各プリンヌクレオチドのリボースの2’位がヒドロキシ基である、アプタマー;
(b)該(a)のアプタマーにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位のフッ素原子が、それぞれ独立して、無置換であるか、水素原子、ヒドロキシ基及びメトキシ基からなる群より選ばれる原子又は基で置換されており、
(ii)各プリンヌクレオチドのリボースの2’位のヒドロキシ基が、それぞれ独立して、無置換であるか、水素原子、メトキシ基及びフッ素原子からなる群より選ばれる原子又は基で置換されている、アプタマー。
[2]N1は、G、GG、AG、C又はギャップ、
N2は、A又はU、
N3は、G、C又はA、
N4は、G、C又はU、
N5は、G又はU、
N6は、UUCN61又はAGUCN62
N61及びN62は、それぞれ独立して任意の0から数個の塩基である、
[1]記載のアプタマー。
[3]下式(2)又は(3)で表わされるヌクレオチド配列を含む、[1]又は[2]記載のアプタマー:
GGGAAACUAGGGCGUUAACGUGACCAGUGUUUCN61(式2)
N1GGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCN62(式3)。
[4]配列番号2又は7で表わされるヌクレオチド配列を含む、[1]~[3]のいずれかに記載のアプタマー。
[5]配列番号1、3、4、5、6、8、10又は11で表わされるヌクレオチド配列を含む、[1]~[3]のいずれかに記載のアプタマー。
[6][1]~[5]のいずれかに記載のアプタマーにおいて、1又は数個のヌクレオチドが置換、欠失、挿入又は付加されたヌクレオチド配列を含むアプタマーであって、
(a)該アプタマーに含まれるヌクレオチドにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位がフッ素原子であり、
(ii)各プリンヌクレオチドのリボースの2’位がヒドロキシ基である、アプタマー;
(b)該(a)のアプタマーにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位のフッ素原子が、それぞれ独立して、無置換であるか、水素原子、ヒドロキシ基及びメトキシ基からなる群より選ばれる原子又は基で置換されており、
(ii)各プリンヌクレオチドのリボースの2’位のヒドロキシ基が、それぞれ独立して、無置換であるか、水素原子、メトキシ基及びフッ素原子からなる群より選ばれる原子又は基で置換されている、アプタマー。
[7]ヌクレオチドの長さが45ヌクレオチド以下である、[1]~[6]のいずれかに記載のアプタマー。
[8]FGF2とFGF受容体との結合を阻害する、[1]~[7]のいずれかに記載のアプタマー。
[9]少なくとも1つのヌクレオチドが修飾されている、[1]~[8]のいずれかに記載のアプタマー。
[10][1]~[9]のいずれかに記載のアプタマー及び機能性物質を含む複合体。
[11]機能性物質が、親和性物質、標識用物質、酵素、薬物送達媒体又は薬物である、[10]記載の複合体。
[12][1]~[9]のいずれかに記載のアプタマーあるいは[10]又は[11]に記載の複合体を含む、医薬。
[13][1]~[9]のいずれかに記載のアプタマーあるいは[10]又は[11]に記載の複合体を含む、血管新生を伴う疾患、骨・軟骨疾患又は疼痛の治療用又は予防用医薬。
[14]血管新生を伴う疾患、骨・軟骨疾患又は疼痛を治療又は予防する方法であって、有効量の[1]~[9]のいずれかに記載のアプタマーあるいは[10]又は[11]に記載の複合体を、対象に投与することを含む方法。
[15]血管新生を伴う疾患、骨・軟骨疾患又は疼痛の治療又は予防に用いるための、[1]~[9]のいずれかに記載のアプタマーあるいは[10]又は[11]に記載の複合体。
[16]血管新生を伴う疾患、骨・軟骨疾患又は疼痛の治療用又は予防用医薬を製造するための、[1]~[9]のいずれかに記載のアプタマーあるいは[10]又は[11]に記載の複合体の使用。
下式(1)
N1GGAN2ACUAGGGCN3UUAAN4GUN5ACCAGUGUN6(式1)
N1及びN6は、それぞれ独立して任意の0から数個の塩基、
N2、N3、N4及びN5は、独立して任意の一個の塩基を表す、
で表わされるヌクレオチド配列(但し、ウラシルはチミンであってもよい)を含むアプタマーであって、以下(a)又は(b)
(a)該アプタマーに含まれるヌクレオチドにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位がフッ素原子であり、
(ii)各プリンヌクレオチドのリボースの2’位がヒドロキシ基である、アプタマー;あるいは
(b)該(a)のアプタマーにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位のフッ素原子が、それぞれ独立して、無置換であるか、水素原子、ヒドロキシ基及びメトキシ基からなる群より選ばれる原子又は基で置換されており、
(ii)各プリンヌクレオチドのリボースの2’位のヒドロキシ基が、それぞれ独立して、無置換であるか、水素原子、メトキシ基及びフッ素原子からなる群より選ばれる原子又は基で置換されている、アプタマー
のいずれかである、FGF2に結合するアプタマー:
を提供する。
よって、本発明のアプタマーがFGF2に結合し、FGF2とFGF受容体との結合を阻害した場合、FGF受容体を介した細胞内シグナル伝達経路の活性化に伴う作用、例えば、細胞死の抑制、細胞増殖、osteoprotegerin(OPG)産生の抑制などが阻害され得る。
N1の塩基数は、式(1)で表わされるヌクレオチド配列を含むアプタマーがFGF2に結合する限り特に限定されないが、例えば、0~約10個、0~9個、0~8個、0~7個、0~6個、0~5個、0~4個、0~3個、0~2個などであってよく、好ましくは0~2個であり得る。
N6の塩基数についてもN1と同様に特に限定されないが、例えば、0~約10個、0~9個、0~8個、0~7個、0~6個、0~5個、0~4個、0~3個などであってよく、好ましくは0~10個、3~9個、又は5~8個である。
N1は、G、GG、AG、C又はギャップであり、
N2は、A又はUであり、
N3は、G、C又はAであり、
N4は、G、C又はUであり、
N5は、G又はUであり、
N6は、UUCN61又はAGUCN62(式中、N61及びN62は、それぞれ独立して任意の0から数個の塩基である)である。ここで、N1が「ギャップ」であるとは、式(1)中にN1が存在しないこと、すなわちN1が0個の塩基であることを意味する。
N61の塩基数は特に限定されないが、例えば、0~約10個、0~7個、0~6個、0~5個、0~4個などであってよく、好ましくは0~5個、1~5個、又は2~4個であり得る。
N62の塩基数についても特に限定されないが、例えば、0~約10個、0~7個、0~5個、0~4個、0~3個などであってよく、好ましくは0~5個、0~4個、又は0~3個であり得る。
別の好ましい実施態様において、上記式(1)中、
N1は、G、GG、AG又はギャップであり、
N2は、A又はUであり、
N3は、G又はAであり、
N4は、C又はUであり、
N5は、G又はUであり、
N6は、UUCN61又はAGUCN62(式中、N61及びN62は上記と同義である)である。
GGGAAACUAGGGCGUUAACGUGACCAGUGUUUCN61(式2)
N1GGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCN62(式3)
(式中、N1、N61及びN62は前記と同義である)
で表されるヌクレオチド配列を含み得る。
配列番号1:
GGGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCUCGA、
配列番号2:
GGGAAACUAGGGCGUUAACGUGACCAGUGUUUCUCGA、
配列番号3:
GGGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCCC、
配列番号4:
GGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCC、
配列番号5:
GGGGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCCCC、
配列番号6:
AGGGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCCC、
配列番号7:
GGGAAACUAGGGCGUUAACGUGACCAGUGUUUCCC、
配列番号8:
CGGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCCG、
配列番号9:
CCGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCGG、
配列番号10:
GGGAUACUAGGGCGUUAACGUUACCAGUGUAGUCCC、
配列番号11:
GGGAUACUAGGGCCUUAAGGUUACCAGUGUAGUCCC、
配列番号12:
GGGAUACUAGGGCAUUU AUGUUACCAGUGUAGUCCC。
好ましい一実施態様において、本発明のアプタマーは、配列番号1、3、4、5、6、8、10又は11で表わされるヌクレオチド配列を含む。
別の好ましい実施態様において、本発明のアプタマーは、配列番号2又は7で表わされるヌクレオチド配列を含む。
さらに別の好ましい実施態様において、本発明のアプタマーは、配列番号1、3、4、5又は6で表わされるヌクレオチド配列を含む。
(a)該アプタマーに含まれるヌクレオチドにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位がフッ素原子であり、
(ii)各プリンヌクレオチドのリボースの2’位がヒドロキシ基である、アプタマー;
(b)該(a)のアプタマーにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位のフッ素原子が、それぞれ独立して、無置換であるか、水素原子、ヒドロキシ基及びメトキシ基からなる群より選ばれる原子又は基で置換されており、
(ii)各プリンヌクレオチドのリボースの2’位のヒドロキシ基が、それぞれ独立して、無置換であるか、水素原子、メトキシ基及びフッ素原子からなる群より選ばれる原子又は基で置換されている、アプタマー
であってよい。ここで、上記置換、欠失、挿入又は付加されるヌクレオチド数は、置換、欠失、挿入又は付加後も依然としてFGF2に結合する限り特に限定されないが、例えば1~約10個、好ましくは1~6個、より好ましくは1~5個、さらに好ましくは1~4個、さらに好ましくは1~3個、最も好ましくは1個又は2個であり得る。ヌクレオチドが置換、欠失、挿入又は付加される部位も、置換、欠失、挿入又は付加後も依然としてFGF2に結合する限り特に限定されないが、上記式(1)、(2)及び(3)中、1種のヌクレオチドに特定されている部位においては、1~3か所、好ましくは1又は2か所、より好ましくは1か所においてヌクレオチドが置換、欠失、挿入又は付加される。一方、式(1)、(2)及び(3)中、複数種のヌクレオチドをとり得る部位においては、より多くのヌクレオチド(例えば、1~約10個、好ましくは1~6個、より好ましくは1~5個、さらに好ましくは1~4個)の置換、欠失、挿入又は付加も許容され得る。
よって本発明のアプタマーの長さとしては、通常約10~約200ヌクレオチドであり得、好ましくは20~80ヌクレオチドであり、より好ましくは25~60ヌクレオチドであり、さらに好ましくは25~50ヌクレオチドであり、最も好ましくは30~45ヌクレオチドである。
ここで連結はタンデム結合にて行われ得る。また、連結に際し、リンカーを利用してもよい。リンカーとしては、ヌクレオチド鎖(例、1~約20ヌクレオチド)、非ヌクレオチド鎖(例、-(CH2)n-リンカー、-(CH2CH2O)n-リンカー、ヘキサエチレングリコールリンカー、TEGリンカー、ペプチドを含むリンカー、-S-S-結合を含むリンカー、-CONH-結合を含むリンカー、-OPO3-結合を含むリンカー)が挙げられる。上記複数の連結物における複数とは、2以上であれば特に限定されないが、例えば2個、3個又は4個であり得る。
本発明のアプタマーにおいてはまた、全てのプリンヌクレオチドが、リボースの2’位のヒドロキシ基が上述した任意の原子又は基、例えば、水素原子、フッ素原子及び-O-Me基からなる群より選ばれる同一の原子又は基で置換されているヌクレオチドであり得る。
本発明のアプタマーにおいて、ウラシルをチミンに置換することによって、FGF2に対する結合性、FGF2とFGF受容体との結合阻害活性、アプタマーの安定性、薬物送達性、血液中での安定性等を高めることが可能である。
連結基としては、-O-、-N-又は-S-が例示され、これらの連結基を通じて隣接するヌクレオチドに結合し得る。
改変はまた、キャッピングのような3’及び5’の改変を含んでもよい。
このようなPEGとしては特に限定されず、当業者であれば市販あるいは公知のPEGを適宜選択して用いることができる(例えば、http://www.peg-drug.com/peg_product/branched.htmlを参照のこと)が、本発明のアプタマーに適用するPEGの好適例として具体的には、分子量40000の2分岐GS型PEG(SUNBRIGHT GL2-400GS 日油製)、分子量40000の2分岐TS型PEG(SUNBRIGHT GL2-400TS 日油製)、分子量40000の4分岐TS型PEG(SUNBRIGHT GL4-400TS 日油製)、分子量80000の2分岐TS型PEG(SUNBRIGHT GL2-800TS 日油製)、又は分子量80000の4分岐TS型PEG(SUNBRIGHT GL4-800TS 日油製)などが挙げられる。
SELEXで得られるアプタマーはそのプライマー設計に依存して、その後の最小化作業のしやすさが変わる。うまくプライマーを設計しないと、SELEXによって活性のあるアプタマーが選別できたとしても、その後の開発が不可能となる。
(i)各ピリミジンヌクレオチドのリボースの2’位がフッ素原子であり、
(ii)各プリンヌクレオチドのリボースの2’位がヒドロキシ基である、アプタマー;(b’)配列番号1~7のいずれか(あるいは、配列番号2もしくは7、又は配列番号1及び3~6のいずれか)で表わされるヌクレオチド配列(但し、ウラシルはチミンであってもよい)において、1~5個(あるいは、1~4個、又は1~3個)のヌクレオチドが置換、欠失、挿入又は付加されたヌクレオチド配列を含むアプタマーであって、該アプタマーに含まれるヌクレオチドにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位フッ素原子であり、
(ii)各プリンヌクレオチドのリボースの2’位がヒドロキシ基である、アプタマー;又は
(c’)該(a’)又は(b’)のアプタマーにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位のフッ素原子が、それぞれ独立して、無置換であるか、水素原子、ヒドロキシ基及びメトキシ基からなる群より選ばれる原子又は基で置換されており、
(ii)各プリンヌクレオチドのリボースの2’位のヒドロキシ基が、それぞれ独立して、無置換であるか、水素原子、メトキシ基及びフッ素原子からなる群より選ばれる原子又は基で置換されている、アプタマー、
であり、さらに好ましくは、上記アプタマーのうちヌクレオチド長が30~45ヌクレオチドであるアプタマーである。
油としては、例えばトウモロコシ油、オリーブ油、綿実油、ヒマワリ油等が挙げられる。また、軟膏剤の場合、適当な医薬上許容される基剤(黄色ワセリン、白色ワセリン、パラフィン、プラスチベース、シリコーン、白色軟膏、ミツロウ、豚油、植物油、親水軟膏、親水ワセリン、精製ラノリン、加水ラノリン、吸水軟膏、親水プラスチベース、マクロゴール軟膏等)を用い、有効成分と混合し製剤化し使用する。
本発明のアプタマーは1本鎖の核酸であるため、相補配列を含むヌクレオチドの投与による解毒も可能であり、投与後の動態制御が困難な中和抗体より安全性の高い医薬品となる可能性が高い。これは、抗体医薬治療などで起こりうる、体内における抗体の長い滞留時間に起因する感染症の問題を鑑みても極めて有利な点と言える。特に本発明の医薬を上記疾患の予防又は治療用医薬として用いる場合、疾患の重篤性と副作用のリスクとを考えると、体内動態を制御しやすいアプタマーを利用する方がより高い安全性を有する医薬を得られることは明白である。
本発明の精製及び濃縮方法はさらに、FGF2の吸着後、洗浄液を用いて当該固相担体を洗浄することを含み得る。洗浄液としては、例えば、尿素、キレート剤(例、EDTA)、Tris、酸、アルカリ、Transfer RNA、DNA、Tween(ツイーン)20などの表面活性剤、NaClなどの塩を含むものなどが挙げられる。本発明の精製及び濃縮方法はさらに、当該固相担体を加熱処理することを含み得る。かかる工程により、当該固相担体の再生、滅菌が可能である。
従来のSELEX法では約30mer~40merのランダム配列の両末端に20mer前後のプライマーを付けたライブラリーが使用された。その場合、取得されるアプタマーの全長は約80~100merであり、その後短鎖化の作業が必要であった。しかし、短鎖化は必ずしも簡単ではなく、活性が大きく低下してしまうことがしばしばであった。そこで、NOXXON社によって開発されたTailored-SELEX法を参考にして(Vater et al.Nucleic Acids Res. 31, 2003,el30; Jarosch et al. Nucleic Acids Res. 34, 2006, e86)、プライマー配列が入らない30mer程度の長さのRNAプールを用いたSELEXを実施した。
使用したDNA鋳型とプライマー配列は以下の通りである。
DNA鋳型:
5’-TCGAG-30N-TCCCTATAGTGAGTCGTATTAGCAGCTCCACAGGCTT-3’(配列番号13)
Forward ligate:
5’-UAAUACGACUCACUAUA-3’(配列番号14)
Forward primer:
5’-AAGCCTGTGGAGCTGCTAATACGACTCACTATAGGGA-3’(配列番号15)
Forward bridge:
5’-TCCCTATAGTGAGTCGTATTA-NH2-3’(配列番号16)
Reverse bridge:
5’-TCTTGTTCAGCTTAGTTCTCTCGAG-3’(配列番号17)
Reverse ligate:
5’-p-GAGAACTAAGCTGAACAAGA-NH2-3’(配列番号18)
アプタマーID1:
GGGAU(F)AC(F)U(F)AGGGC(F)AU(F)U(F)AAU(F)GU(F)U(F)AC(F)C(F)AGU(F)GU(F)AGU(F)C(F)U(F)C(F)GA
アプタマーID2:
GGGAAAC(F)U(F)AGGGC(F)GU(F)U(F)AAC(F)GU(F)GAC(F)C(F)AGU(F)GU(F)U(F)U(F)C(F)U(F)C(F)GA
配列番号1及び2で表わされるアプタマーの短鎖化を行った。MFOLDプログラム(Zuker,Nucleic Acids Res.31,3406-3415,2003)を用いてRNAの2次構造を予測し、その構造を参考に短鎖化した。短鎖化体は、目的の配列のDNAを化学合成により作製し、DuraScribe T7 Transcription Kitを用いて転写することで得られた。以下に、実際に作製した短鎖化体のヌクレオチド配列(配列番号3及び7)をリボースの2’位の修飾と共にアプタマーID3及び7として示す。
GGGAU(F)AC(F)U(F)AGGGC(F)AU(F)U(F)AAU(F)GU(F)U(F)AC(F)C(F)AGU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID7:配列番号2で表わされるアプタマー改変体で35ヌクレオチドの長さのアプタマー
GGGAAAC(F)U(F)AGGGC(F)GU(F)U(F)AAC(F)GU(F)GAC(F)C(F)AGU(F)GU(F)U(F)U(F)C(F)C(F)C(F)
アプタマーID3で表わされるFGF2アプタマーが同じFGFファミリーのFGF1、又はいくつかの成長因子EGF、β―NGF、VEGFに結合するかどうか表面プラズモン共鳴法で調べた。測定にはBIAcore社製のBIAcore2000を用いた。センサーチップにはストレプトアビジンが固定化されているSAチップを用いた。これに、5末端にビオチンが付加したアプタマーID3で表わされるアプタマーを約500RU程度結合させた。ビオチン付きアプタマーは化学合成によって作製した。リガンドとなるタンパク質はR&D社製のFGF1、EGF、β―NGF、VEGFを用いた。ランニングバッファーは実施例1で使用した溶液Aに最終濃度0.3Mになるように塩化ナトリウムを添加したものを用いた。その結果、アプタマーID3で表わされるアプタマーはFGF2とは結合するが、他のタンパク質とは結合しないことがわかった。そのセンサーグラムを図3に示す。
以上より、アプタマーID3で表わされるアプタマーはFGF2に特異的に結合することが判明した。
FGF2に対する結合性、安定性、薬物送達性などを高めるため、配列番号3で表わされるアプタマーを基にして、アプタマーID3(1)-3(40)、アプタマーID4及び4(1)―4(4)、アプタマーID5並びにアプタマーID6で表わされる核酸を化学合成した。ここで、アプタマーID4で表わされるアプタマーはアプタマーID3(19)で表わされるアプタマーの5’末端のG(M)及び3’末端のC(M)をそれぞれ一つ削ったものであった。アプタマーID5で表わされるアプタマーはアプタマーID3(19)で表わされるアプタマーの5’末端にG(M)及び3’末端にC(M)をそれぞれ一つ追加したものであった。また、アプタマーID6で表わされるアプタマーはアプタマーID3(19)で表わされるアプタマーの5’末端にA(M)だけを追加したものであった。これらの核酸は化学合成により作製した。作製したアプタマーがFGF2とFGF受容体の結合を阻害するかどうか実施例1と同様に調べた。ここでは、アプタマー、FGF2、ヘパリンの濃度をすべて0.1μMとした。実験の結果、測定した全てのアプタマーがFGF2とFGFR1α(IIIc)受容体の結合を強く阻害することがわかった。表3にその結果を示す。
GGGAU(F)AC(F)U(F)AGGGC(F)A(M)U(F)U(F)A(M)A(M)U(F)G(M)U(F)U(F)AC(F)C(F)AGU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(2)
G(M)G(M)G(M)A(M)U(F)AC(F)U(F)AGGGC(F)AU(F)U(F)AAU(F)GU(F)U(F)AC(F)C(F)AGU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(3)
GGGAU(F)AC(F)U(F)AGG(M)GC(F)AU(F)U(F)AAU(F)GU(F)U(F)AC(F)C(F)AGU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(4)
GGGAU(F)AC(F)U(F)AG(M)GGC(F)AU(F)U(F)AAU(F)GU(F)U(F)AC(F)C(F)AGU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(5)
GGGAU(F)AC(F)U(F)A(M)GGGC(F)AU(F)U(F)AAU(F)GU(F)U(F)AC(F)C(F)AGU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(6)
GGGAU(F)AC(F)U(F)AGGGC(F)AU(F)U(F)AAU(F)GU(F)U(F)A(M)C(F)C(F)AGU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(7)
GGGAU(F)AC(F)U(F)AGGGC(F)AU(F)U(F)AAU(F)GU(F)U(F)AC(F)C(F)A(M)GU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(8)
G(M)G(M)G(M)A(M)U(F)AC(F)U(F)A(M)G(M)G(M)GC(F)A(M)U(F)U(F)A(M)A(M)U(F)G(M)U(F)U(F)A(M)C(F)C(F)A(M)GU(F)GU(F)AGU(F)C(F)C(F))C(F)-idT
アプタマーID3(9)
GGGAU(F)A(M)C(F)U(F)AGGGC(F)AU(F)U(F)AAU(F)GU(F)U(F)AC(F)C(F)AGU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(10)
GGGAU(F)AC(F)U(F)AGGGC(F)AU(F)U(F)AAU(F)GU(F)U(F)AC(F)C(F)AGU(F)GU(F)AG(M)U(F)C(F)C(F)C(F)
アプタマーID3(11)
GGGAU(F)AC(F)U(F)AGGGC(F)AU(F)U(F)AAU(F)GU(F)U(F)AC(F)C(F)AGU(F)GU(F)A(M)GU(F)C(F)C(F)C(F)
アプタマーID3(12)
G(M)G(M)G(M)A(M)U(F)AC(F)U(F)A(M)G(M)G(M)GC(F)A(M)U(F)U(F)A(M)A(M)U(F)G(M)U(F)U(F)A(M)C(F)C(F)A(M)GU(F)GU(F)AGU(M)C(M)C(M)C(M)-idT
アプタマーID3(13)
G(M)G(M)G(M)A(M)U(F)AC(F)U(F)A(M)G(M)G(M)GC(F)A(M)U(M)U(M)A(M)A(M)U(M)G(M)U(F)U(F)A(M)C(F)C(F)A(M)GU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(14)
G(M)G(M)G(M)A(M)U(F)AC(F)U(F)A(M)G(M)G(M)GC(F)A(M)U(F)U(F)A(M)A(M)U(F)G(M)U(F)U(M)A(M)C(F)C(F)A(M)GU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(15)
G(M)G(M)G(M)A(M)U(F)AC(F)U(F)A(M)G(M)G(M)GC(F)A(M)U(F)U(F)A(M)A(M)U(F)G(M)U(F)U(F)A(M)C(M)C(M)A(M)GU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(16)
G(M)G(M)G(M)A(M)U(F)AC(M)U(F)A(M)G(M)G(M)GC(F)A(M)U(F)U(F)A(M)A(M)U(F)G(M)U(F)U(F)A(M)C(F)C(F)A(M)GU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(17)
G(M)G(M)G(M)A(M)U(M)AC(F)U(F)A(M)G(M)G(M)GC(F)A(M)U(F)U(F)A(M)A(M)U(F)G(M)U(F)U(F)A(M)C(F)C(F)A(M)GU(F)GU(F)AGU(F)C(F)C(F)C(F)
アプタマーID3(18)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(19)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)-idT
アプタマーID3(20)
idT-G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(21)
GL2-400TS-C6-G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)-idT
アプタマーID3(22)
idT-G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)-C6-GL2-400TS
アプタマーID3(23)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)gC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(24)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)gU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(25)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)G(F)C(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(26)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)G(F)U(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(27)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)G(F)U(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(28)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)sGC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(29)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GsC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(30)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)sGU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(31)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GsU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(32)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)sGU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(33)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GsU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID3(34)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)G(F)C(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)G(F)U(F)G(F)U(F)A(M)G(M)U(M)C(M)C(M)C(M)-idT
アプタマーID3(35)
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)G(F)C(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)G(F)U(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)-idT
アプタマーID3(36)
GL4-800TS-C6-G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)-idT
アプタマーID3(37)
Y-NHS-40K-ssH-G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)-idT
アプタマーID3(38)
ME-100TS-C6-G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)-idT
アプタマーID3(39)
PTE-100CS-C6-G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)-idT
アプタマーID3(40)
GL2-400TS-ssH-G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)-idT
アプタマーID4:アプタマーID3(19)で表わされるアプタマーの改変体で34ヌクレオチドの長さのアプタマー
G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)-idT
アプタマーID4(1)
G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)G(F)C(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)-idT
アプタマーID4(2)
G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)G(F)C(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)G(F)U(F)GU(F)A(M)G(M)U(M)C(M)C(M)-idT
アプタマーID4(3)
G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)G(F)C(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)sGU(F)A(M)G(M)U(M)C(M)C(M)-idT
アプタマーID4(4)
G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)G(F)C(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)G(F)U(F)GsU(F)A(M)G(M)U(M)C(M)C(M)-idT
アプタマーID5:アプタマーID3(19)で表わされるアプタマーの改変体で38ヌクレオチドの長さのアプタマー
G(M)G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)C(M)-idT
アプタマーID6:アプタマーID3(19)で表わされるアプタマーの改変体で37ヌクレオチドの長さのアプタマー
A(M)G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)-idT
ヒト臍帯静脈血管内皮細胞(HUVEC)を96ウェル平底プレートに1ウェル当たり5x103個播種し、2%ウシ胎児血清と成長因子を含んだ内皮細胞専用培地EGM-2 BulletKit(CC-3162 Lonza社製)で一晩培養した。その後、培地を捨て、PBSバッファーで2回洗浄後、2%ウシ胎児血清を含んだ内皮細胞専用培地に溶解したアプタマーID3(21)で表わされるアプタマー(5、2.5、1、0.5nM)とFGF2(最終濃度0.5nM)の混合液を添加した。72時間後にCell Counting Kit-8を用いて生細胞数を調べた。吸光度測定にはマイクロプレートリーダー(450nm)を使用した。1サンプルはn=3で測定した。ポジティブコントロールとして抗FGF2抗体Anti-FGF, basic(Ab-3) Mouse mAb(3H3)(Calbiochem社製)を用いた。FGF2のみの添加で細胞を3日間培養して得られた1ウェルあたりのOD値を阻害活性0%、FGF2無添加で3日間培養して得られた1ウェルあたりのOD値を阻害活性100%として、FGF2とアプタマーの混合液を添加した場合に得られた1ウェルあたりのOD値から、アプタマーの阻害活性を求めた。その結果、アプタマーID3(21)で表わされるアプタマーがFGF2に対して高い阻害活性を有していることが示された。IC50値は約1.0nMであった。結果を表4に示す。
実施例5と同様な方法で、アプタマーID8から12で表されるアプタマーの阻害活性を測定した。その結果を表6に示す。
アプタマーID8~12のヌクレオチド配列はそれぞれ配列番号8~12で表される。
NH2-C(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)G(M)-idT
アプタマーID9
NH2-C(M)C(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)A(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)G(M)G(M)-idT
アプタマーID10
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)G(M)U(M)U(F)A(M)A(M)C(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID11
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)C(M)U(M)U(F)A(M)A(M)G(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
アプタマーID12
G(M)G(M)G(M)A(M)U(M)A(M)C(M)U(F)A(M)G(M)G(M)GC(M)A(M)U(M)U(F)U(M)A(M)U(M)G(M)U(F)U(M)A(M)C(M)C(M)A(M)GU(F)GU(F)A(M)G(M)U(M)C(M)C(M)C(M)
ヒトFGF-2(R&D社製)を含む0.76%(最終濃度)のクエン酸ナトリウム含有マトリゲル(BDマトリゲルTM)を8週令のC57BL/6Jマウス(♀)の皮下に麻酔下で注入した。7日後にマトリゲルを摘出し、血管新生の度合いをマトリゲル中のヘモグロビン量で評価した。ヘモグロビン量はDrabkin Reagent Kitを用いてシアンメトヘモグロビン法で定量した。アプタマーは1mM塩化マグネシウムを含んだリン酸緩衝液に溶解し、マトリゲル皮下投与直後から、一日1回腹腔内に投与した。投与群を表7に、結果を表8に示す。アプタマー1mg/kg群で顕著な血管新生阻害が観察された。以上より、本発明のアプタマーがモデル動物においても強い血管新生阻害活性を示すことが確認された。
Claims (16)
- 下式(1)で表わされるヌクレオチド配列(但し、ウラシルはチミンであってもよい)を含むアプタマーであって、以下(a)又は(b)のいずれかである、FGF2に結合するアプタマー:
N1GGAN2ACUAGGGCN3UUAAN4GUN5ACCAGUGUN6(式1)
N1及びN6は、それぞれ独立して任意の0から数個の塩基、
N2、N3、N4及びN5は、独立して任意の一個の塩基を表す、
(a)該アプタマーに含まれるヌクレオチドにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位がフッ素原子であり、
(ii)各プリンヌクレオチドのリボースの2’位がヒドロキシ基である、アプタマー;
(b)該(a)のアプタマーにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位のフッ素原子が、それぞれ独立して、無置換であるか、水素原子、ヒドロキシ基及びメトキシ基からなる群より選ばれる原子又は基で置換されており、
(ii)各プリンヌクレオチドのリボースの2’位のヒドロキシ基が、それぞれ独立して、無置換であるか、水素原子、メトキシ基及びフッ素原子からなる群より選ばれる原子又は基で置換されている、アプタマー。 - N1は、G、GG、AG、C又はギャップ、
N2は、A又はU、
N3は、G、C又はA、
N4は、G、C又はU、
N5は、G又はU、
N6は、UUCN61又はAGUCN62
N61及びN62は、それぞれ独立して任意の0から数個の塩基である、
請求項1記載のアプタマー。 - 下式(2)又は(3)で表わされるヌクレオチド配列を含む、請求項1又は2記載のアプタマー:
GGGAAACUAGGGCGUUAACGUGACCAGUGUUUCN61(式2)
N1GGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCN62(式3)。 - 配列番号2又は7で表わされるヌクレオチド配列を含む、請求項1~3のいずれか一項に記載のアプタマー。
- 配列番号1、3、4、5、6、8、10又は11で表わされるヌクレオチド配列を含む、請求項1~3のいずれか一項に記載のアプタマー。
- 請求項1~5のいずれか一項に記載のアプタマーにおいて、1又は数個のヌクレオチドが置換、欠失、挿入又は付加されたヌクレオチド配列を含むアプタマーであって、
(a)該アプタマーに含まれるヌクレオチドにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位がフッ素原子であり、
(ii)各プリンヌクレオチドのリボースの2’位がヒドロキシ基である、アプタマー;
(b)該(a)のアプタマーにおいて、
(i)各ピリミジンヌクレオチドのリボースの2’位のフッ素原子が、それぞれ独立して、無置換であるか、水素原子、ヒドロキシ基及びメトキシ基からなる群より選ばれる原子又は基で置換されており、
(ii)各プリンヌクレオチドのリボースの2’位のヒドロキシ基が、それぞれ独立して、無置換であるか、水素原子、メトキシ基及びフッ素原子からなる群より選ばれる原子又は基で置換されている、アプタマー。 - ヌクレオチドの長さが45ヌクレオチド以下である、請求項1~6のいずれか一項に記載のアプタマー。
- FGF2とFGF受容体との結合を阻害する、請求項1~7のいずれか一項に記載のアプタマー。
- 少なくとも1つのヌクレオチドが修飾されている、請求項1~8のいずれか一項に記載のアプタマー。
- 請求項1~9のいずれか一項に記載のアプタマー及び機能性物質を含む複合体。
- 機能性物質が、親和性物質、標識用物質、酵素、薬物送達媒体又は薬物である、請求項10記載の複合体。
- 請求項1~9のいずれか一項に記載のアプタマーあるいは請求項10又は11に記載の複合体を含む、医薬。
- 請求項1~9のいずれか一項に記載のアプタマーあるいは請求項10又は11に記載の複合体を含む、血管新生を伴う疾患、骨・軟骨疾患又は疼痛の治療用又は予防用医薬。
- 血管新生を伴う疾患、骨・軟骨疾患又は疼痛を治療又は予防する方法であって、有効量の請求項1~9のいずれか一項に記載のアプタマーあるいは請求項10又は11に記載の複合体を、対象に投与することを含む方法。
- 血管新生を伴う疾患、骨・軟骨疾患又は疼痛の治療又は予防に用いるための、請求項1~9のいずれか一項に記載のアプタマーあるいは請求項10又は11に記載の複合体。
- 血管新生を伴う疾患、骨・軟骨疾患又は疼痛の治療用又は予防用医薬を製造するための、請求項1~9のいずれか一項に記載のアプタマーあるいは請求項10又は11に記載の複合体の使用。
Priority Applications (15)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US15/128,495 US9987298B2 (en) | 2014-03-24 | 2015-03-24 | Aptamer for FGF2 and use thereof |
KR1020167029463A KR102394676B1 (ko) | 2014-03-24 | 2015-03-24 | Fgf2에 대한 압타머 및 그의 사용 |
EP15767725.3A EP3124611B1 (en) | 2014-03-24 | 2015-03-24 | Aptamer for fgf2 and use thereof |
JP2016510402A JP6634554B2 (ja) | 2014-03-24 | 2015-03-24 | Fgf2に対するアプタマー及びその使用 |
CA2943772A CA2943772C (en) | 2014-03-24 | 2015-03-24 | Aptamer for fgf2 and use thereof |
RU2016141263A RU2686992C2 (ru) | 2014-03-24 | 2015-03-24 | Аптамер для fgf2 и его применение |
PL15767725T PL3124611T3 (pl) | 2014-03-24 | 2015-03-24 | Aptamer dla FGF2 i jego zastosowanie |
SG11201607906RA SG11201607906RA (en) | 2014-03-24 | 2015-03-24 | Aptamer for fgf2 and use thereof |
DK15767725.3T DK3124611T3 (da) | 2014-03-24 | 2015-03-24 | Aptamer til fgf2 og anvendelse deraf |
AU2015235020A AU2015235020B2 (en) | 2014-03-24 | 2015-03-24 | Aptamer for FGF2 and use thereof |
ES15767725T ES2768693T3 (es) | 2014-03-24 | 2015-03-24 | Aptámero para FGF2 y uso del mismo |
MX2016012466A MX2016012466A (es) | 2014-03-24 | 2015-03-24 | Aptamero para el factor de crecimiento de fibroblastos 2 y uso del mismo. |
BR112016020984A BR112016020984A2 (pt) | 2014-03-24 | 2015-03-24 | Aptâmero para fgf2 e uso do mesmo |
CN201580015642.1A CN106103719B (zh) | 2014-03-24 | 2015-03-24 | 针对fgf2的适体及其应用 |
IL247998A IL247998B (en) | 2014-03-24 | 2016-09-22 | aptamer for fgf2 and used in it |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2014060966 | 2014-03-24 | ||
JP2014-060966 | 2014-03-24 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2015147017A1 true WO2015147017A1 (ja) | 2015-10-01 |
Family
ID=54195529
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2015/058992 WO2015147017A1 (ja) | 2014-03-24 | 2015-03-24 | Fgf2に対するアプタマー及びその使用 |
Country Status (18)
Country | Link |
---|---|
US (1) | US9987298B2 (ja) |
EP (1) | EP3124611B1 (ja) |
JP (1) | JP6634554B2 (ja) |
KR (1) | KR102394676B1 (ja) |
CN (1) | CN106103719B (ja) |
AU (1) | AU2015235020B2 (ja) |
BR (1) | BR112016020984A2 (ja) |
CA (1) | CA2943772C (ja) |
DK (1) | DK3124611T3 (ja) |
ES (1) | ES2768693T3 (ja) |
HU (1) | HUE048529T2 (ja) |
IL (1) | IL247998B (ja) |
MX (1) | MX2016012466A (ja) |
PL (1) | PL3124611T3 (ja) |
PT (1) | PT3124611T (ja) |
RU (1) | RU2686992C2 (ja) |
SG (1) | SG11201607906RA (ja) |
WO (1) | WO2015147017A1 (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020004607A1 (ja) | 2018-06-29 | 2020-01-02 | 株式会社リボミック | アプタマー製剤 |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114853869B (zh) * | 2019-12-10 | 2023-12-26 | 湖南赛奥维生物技术有限公司 | 一种碱性成纤维细胞生长因子替代物及其组合物和应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008206524A (ja) * | 1992-09-29 | 2008-09-11 | Gilead Sciences Inc | 核酸リガンドおよびその製造方法 |
WO2011099576A1 (ja) * | 2010-02-12 | 2011-08-18 | 国立大学法人 東京大学 | Fgf2に対するアプタマー及びその使用 |
WO2013186857A1 (ja) * | 2012-06-12 | 2013-12-19 | 株式会社リボミック | Fgf2に対するアプタマー及びその使用 |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5475096A (en) | 1990-06-11 | 1995-12-12 | University Research Corporation | Nucleic acid ligands |
US6177557B1 (en) * | 1990-06-11 | 2001-01-23 | Nexstar Pharmaceuticals, Inc. | High affinity ligands of basic fibroblast growth factor and thrombin |
ES2257735T3 (es) | 1992-09-29 | 2006-08-01 | Gilead Sciences, Inc. | Ligandos de acido nucleico y procedimiento de produccion. |
WO1995007364A1 (en) | 1993-09-08 | 1995-03-16 | Nexstar Pharmaceuticals, Inc. | Nucleic acid ligands and improved methods for producing the same |
US7795419B2 (en) * | 2004-05-26 | 2010-09-14 | Rosetta Genomics Ltd. | Viral and viral associated miRNAs and uses thereof |
RU2410432C1 (ru) * | 2009-11-23 | 2011-01-27 | Государственное учреждение Научно-исследовательский институт физико-химической биологии имени А.Н. Белозерского Московского государственного университета имени М.В. Ломоносова | Модифицированные днк аптамеры, ингибирующие активность тромбина |
SG184497A1 (en) * | 2010-04-12 | 2012-11-29 | Somalogic Inc | 5-position modified pyrimidines and their use |
-
2015
- 2015-03-24 WO PCT/JP2015/058992 patent/WO2015147017A1/ja active Application Filing
- 2015-03-24 CN CN201580015642.1A patent/CN106103719B/zh active Active
- 2015-03-24 PL PL15767725T patent/PL3124611T3/pl unknown
- 2015-03-24 AU AU2015235020A patent/AU2015235020B2/en active Active
- 2015-03-24 BR BR112016020984A patent/BR112016020984A2/pt not_active IP Right Cessation
- 2015-03-24 US US15/128,495 patent/US9987298B2/en active Active
- 2015-03-24 MX MX2016012466A patent/MX2016012466A/es active IP Right Grant
- 2015-03-24 CA CA2943772A patent/CA2943772C/en active Active
- 2015-03-24 JP JP2016510402A patent/JP6634554B2/ja active Active
- 2015-03-24 RU RU2016141263A patent/RU2686992C2/ru active
- 2015-03-24 PT PT157677253T patent/PT3124611T/pt unknown
- 2015-03-24 ES ES15767725T patent/ES2768693T3/es active Active
- 2015-03-24 DK DK15767725.3T patent/DK3124611T3/da active
- 2015-03-24 KR KR1020167029463A patent/KR102394676B1/ko active IP Right Grant
- 2015-03-24 HU HUE15767725A patent/HUE048529T2/hu unknown
- 2015-03-24 SG SG11201607906RA patent/SG11201607906RA/en unknown
- 2015-03-24 EP EP15767725.3A patent/EP3124611B1/en active Active
-
2016
- 2016-09-22 IL IL247998A patent/IL247998B/en active IP Right Grant
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008206524A (ja) * | 1992-09-29 | 2008-09-11 | Gilead Sciences Inc | 核酸リガンドおよびその製造方法 |
WO2011099576A1 (ja) * | 2010-02-12 | 2011-08-18 | 国立大学法人 東京大学 | Fgf2に対するアプタマー及びその使用 |
WO2013186857A1 (ja) * | 2012-06-12 | 2013-12-19 | 株式会社リボミック | Fgf2に対するアプタマー及びその使用 |
Non-Patent Citations (6)
Title |
---|
CANNONE, J. J. ET AL.: "Crystallization of bFGF- DNA aptamer complexes using a sparse matrix designed for protein-nucleic acid complexes", JOURNAL OF CRYSTAL GROWTH, vol. 232, 2001, pages 409 - 417, XP004300538 * |
ISHIGURO, A. ET AL.: "In vivo analysis of FGF2 in murine model of rheumatoid arthritis by RNA aptamer", 14TH RNA MEETING IN TOHOKU YOSHISHU, 2012, pages 60, O-19, XP008184938 * |
JAROSCH, F. ET AL.: "In vitro selection using a dual RNA library that allows primerless selection", NUCLEIC ACIDS RES., vol. 34, no. 12, e86, 2006, pages 1 - 9, XP002404510 * |
KEEFE, A. D. ET AL.: "SELEX with modified nucleotides", CURR. OPIN. CHEM. BIOL., vol. 12, 2008, pages 448 - 456, XP024528065 * |
See also references of EP3124611A4 * |
VATER, A. ET AL.: "Short bioactive Spiegelmers to migraine-associated calcitonin gene -related peptide rapidly identified by a novel approach: tailored-SELEX", NUCLEIC ACIDS RES., vol. 31, no. 21, e130, 2003, pages 1 - 7, XP002524779 * |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020004607A1 (ja) | 2018-06-29 | 2020-01-02 | 株式会社リボミック | アプタマー製剤 |
KR20210025083A (ko) | 2018-06-29 | 2021-03-08 | 가부시키가이샤 리보믹 | 압타머 제제 |
JPWO2020004607A1 (ja) * | 2018-06-29 | 2021-07-15 | 株式会社リボミック | アプタマー製剤 |
US20210269802A1 (en) * | 2018-06-29 | 2021-09-02 | Ribomic Inc. | Aptamer preparation |
JP7340264B2 (ja) | 2018-06-29 | 2023-09-07 | 株式会社リボミック | アプタマー製剤 |
Also Published As
Publication number | Publication date |
---|---|
PT3124611T (pt) | 2020-02-17 |
RU2016141263A3 (ja) | 2018-10-12 |
EP3124611B1 (en) | 2019-11-06 |
CA2943772A1 (en) | 2015-10-01 |
AU2015235020A1 (en) | 2016-11-10 |
IL247998A0 (en) | 2016-11-30 |
US9987298B2 (en) | 2018-06-05 |
EP3124611A4 (en) | 2017-10-25 |
MX2016012466A (es) | 2017-01-06 |
JPWO2015147017A1 (ja) | 2017-04-13 |
US20170157165A1 (en) | 2017-06-08 |
SG11201607906RA (en) | 2016-11-29 |
HUE048529T2 (hu) | 2020-07-28 |
KR102394676B1 (ko) | 2022-05-09 |
EP3124611A1 (en) | 2017-02-01 |
BR112016020984A2 (pt) | 2017-10-03 |
IL247998B (en) | 2020-09-30 |
CN106103719A (zh) | 2016-11-09 |
RU2016141263A (ru) | 2018-04-26 |
DK3124611T3 (da) | 2020-01-27 |
PL3124611T3 (pl) | 2020-05-18 |
CA2943772C (en) | 2022-07-19 |
JP6634554B2 (ja) | 2020-01-22 |
AU2015235020B2 (en) | 2020-10-22 |
RU2686992C2 (ru) | 2019-05-06 |
KR20160135819A (ko) | 2016-11-28 |
ES2768693T3 (es) | 2020-06-23 |
CN106103719B (zh) | 2019-12-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
KR102021626B1 (ko) | Ngf에 대한 압타머 및 그의 용도 | |
JP5704638B2 (ja) | Il−17に対するアプタマー及びその使用 | |
JP5190573B2 (ja) | ミッドカインに対するアプタマー及びその使用 | |
WO2013186857A1 (ja) | Fgf2に対するアプタマー及びその使用 | |
JP5899550B2 (ja) | Fgf2に対するアプタマー及びその使用 | |
JP5810356B2 (ja) | キマーゼに対するアプタマー及びその使用 | |
WO2011118682A1 (ja) | Ngfに対するアプタマー及びその使用 | |
JP6634554B2 (ja) | Fgf2に対するアプタマー及びその使用 | |
WO2020204151A1 (ja) | Fgf9に対するアプタマー及びその使用 | |
WO2019107532A1 (ja) | キマーゼに対するアプタマー及びその使用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 15767725 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 2016510402 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 247998 Country of ref document: IL |
|
ENP | Entry into the national phase |
Ref document number: 2943772 Country of ref document: CA |
|
WWE | Wipo information: entry into national phase |
Ref document number: 15128495 Country of ref document: US Ref document number: MX/A/2016/012466 Country of ref document: MX |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 20167029463 Country of ref document: KR Kind code of ref document: A |
|
ENP | Entry into the national phase |
Ref document number: 2016141263 Country of ref document: RU Kind code of ref document: A |
|
REEP | Request for entry into the european phase |
Ref document number: 2015767725 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2015767725 Country of ref document: EP |
|
REG | Reference to national code |
Ref country code: BR Ref legal event code: B01A Ref document number: 112016020984 Country of ref document: BR |
|
ENP | Entry into the national phase |
Ref document number: 2015235020 Country of ref document: AU Date of ref document: 20150324 Kind code of ref document: A |
|
ENP | Entry into the national phase |
Ref document number: 112016020984 Country of ref document: BR Kind code of ref document: A2 Effective date: 20160912 |