JP6308540B2 - Photodegradable coupling agent - Google Patents
Photodegradable coupling agent Download PDFInfo
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- JP6308540B2 JP6308540B2 JP2017113211A JP2017113211A JP6308540B2 JP 6308540 B2 JP6308540 B2 JP 6308540B2 JP 2017113211 A JP2017113211 A JP 2017113211A JP 2017113211 A JP2017113211 A JP 2017113211A JP 6308540 B2 JP6308540 B2 JP 6308540B2
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- coupling agent
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- 239000007822 coupling agent Substances 0.000 title claims description 47
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 78
- 239000000758 substrate Substances 0.000 description 78
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 72
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 52
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 45
- 238000006303 photolysis reaction Methods 0.000 description 42
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 36
- -1 2-nitrobenzyl group Chemical group 0.000 description 34
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 34
- 230000015843 photosynthesis, light reaction Effects 0.000 description 29
- 239000000243 solution Substances 0.000 description 26
- 235000002597 Solanum melongena Nutrition 0.000 description 22
- 238000010586 diagram Methods 0.000 description 18
- 238000003786 synthesis reaction Methods 0.000 description 18
- 230000005284 excitation Effects 0.000 description 17
- 229910052757 nitrogen Inorganic materials 0.000 description 17
- 230000015572 biosynthetic process Effects 0.000 description 16
- WEVYAHXRMPXWCK-UHFFFAOYSA-N acetonitrile Substances CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 15
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 14
- 239000012299 nitrogen atmosphere Substances 0.000 description 13
- 239000007787 solid Substances 0.000 description 13
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 12
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 12
- 238000004833 X-ray photoelectron spectroscopy Methods 0.000 description 12
- WYURNTSHIVDZCO-UHFFFAOYSA-N tetrahydrofuran Substances C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 12
- 239000006087 Silane Coupling Agent Substances 0.000 description 11
- 238000004440 column chromatography Methods 0.000 description 11
- 230000008859 change Effects 0.000 description 10
- 230000001678 irradiating effect Effects 0.000 description 10
- YXFVVABEGXRONW-UHFFFAOYSA-N toluene Substances CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 10
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 9
- 239000000203 mixture Substances 0.000 description 9
- 239000010703 silicon Substances 0.000 description 9
- 229910052710 silicon Inorganic materials 0.000 description 9
- 150000001345 alkine derivatives Chemical class 0.000 description 8
- 150000001540 azides Chemical class 0.000 description 8
- 229910000365 copper sulfate Inorganic materials 0.000 description 8
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 8
- 238000005259 measurement Methods 0.000 description 8
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 8
- 229910052753 mercury Inorganic materials 0.000 description 8
- 125000003808 silyl group Chemical group [H][Si]([H])([H])[*] 0.000 description 8
- 238000004506 ultrasonic cleaning Methods 0.000 description 8
- IVRMZWNICZWHMI-UHFFFAOYSA-N azide group Chemical group [N-]=[N+]=[N-] IVRMZWNICZWHMI-UHFFFAOYSA-N 0.000 description 7
- 125000005647 linker group Chemical group 0.000 description 7
- 230000004048 modification Effects 0.000 description 7
- 238000012986 modification Methods 0.000 description 7
- 239000012044 organic layer Substances 0.000 description 7
- 239000011780 sodium chloride Substances 0.000 description 7
- 125000003396 thiol group Chemical class [H]S* 0.000 description 7
- 238000001291 vacuum drying Methods 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- 229910014033 C-OH Inorganic materials 0.000 description 6
- 229910014570 C—OH Inorganic materials 0.000 description 6
- 238000005481 NMR spectroscopy Methods 0.000 description 6
- 125000002729 alkyl fluoride group Chemical group 0.000 description 6
- 125000003277 amino group Chemical group 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 230000008878 coupling Effects 0.000 description 6
- 238000010168 coupling process Methods 0.000 description 6
- 238000005859 coupling reaction Methods 0.000 description 6
- 239000003431 cross linking reagent Substances 0.000 description 6
- 239000013078 crystal Substances 0.000 description 6
- 239000003921 oil Substances 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
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- 150000001412 amines Chemical group 0.000 description 5
- 230000002209 hydrophobic effect Effects 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 125000006239 protecting group Chemical group 0.000 description 5
- 239000000377 silicon dioxide Substances 0.000 description 5
- TVUUWNOFLFDCFQ-UHFFFAOYSA-N 6-bromo-4-(chloromethyl)-7-hydroxychromen-2-one Chemical compound O1C(=O)C=C(CCl)C2=C1C=C(O)C(Br)=C2 TVUUWNOFLFDCFQ-UHFFFAOYSA-N 0.000 description 4
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 4
- 239000012620 biological material Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000011259 mixed solution Substances 0.000 description 4
- 238000000059 patterning Methods 0.000 description 4
- 229920001223 polyethylene glycol Polymers 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- NMJQDXLCXGUCPY-UHFFFAOYSA-N (6-bromo-7-hydroxy-2-oxochromen-4-yl)methyl 5-trimethoxysilylpentanoate Chemical compound C1=C(O)C(Br)=CC2=C1OC(=O)C=C2COC(=O)CCCC[Si](OC)(OC)OC NMJQDXLCXGUCPY-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 150000001299 aldehydes Chemical class 0.000 description 3
- 125000000304 alkynyl group Chemical group 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
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- 239000003814 drug Substances 0.000 description 3
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- 229910010272 inorganic material Inorganic materials 0.000 description 3
- 239000011147 inorganic material Substances 0.000 description 3
- 150000002576 ketones Chemical class 0.000 description 3
- 239000003607 modifier Substances 0.000 description 3
- 125000000018 nitroso group Chemical group N(=O)* 0.000 description 3
- 229920000575 polymersome Polymers 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000013545 self-assembled monolayer Substances 0.000 description 3
- 238000007086 side reaction Methods 0.000 description 3
- 125000006850 spacer group Chemical group 0.000 description 3
- LFQCEHFDDXELDD-UHFFFAOYSA-N tetramethyl orthosilicate Chemical compound CO[Si](OC)(OC)OC LFQCEHFDDXELDD-UHFFFAOYSA-N 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- BSKHPKMHTQYZBB-UHFFFAOYSA-N 2-methylpyridine Chemical compound CC1=CC=CC=N1 BSKHPKMHTQYZBB-UHFFFAOYSA-N 0.000 description 2
- SJECZPVISLOESU-UHFFFAOYSA-N 3-trimethoxysilylpropan-1-amine Chemical compound CO[Si](OC)(OC)CCCN SJECZPVISLOESU-UHFFFAOYSA-N 0.000 description 2
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 2
- WFHWWDOGSNSZLP-UHFFFAOYSA-N 4-ethyl-2-methoxy-1-phenylmethoxybenzene Chemical compound COC1=CC(CC)=CC=C1OCC1=CC=CC=C1 WFHWWDOGSNSZLP-UHFFFAOYSA-N 0.000 description 2
- IIDQAAJTECOGEG-UHFFFAOYSA-N 6-bromo-7-hydroxy-4-(hydroxymethyl)chromen-2-one Chemical compound C1=C(O)C(Br)=CC2=C1OC(=O)C=C2CO IIDQAAJTECOGEG-UHFFFAOYSA-N 0.000 description 2
- FRCHGJBAEMHJOF-UHFFFAOYSA-N C(C1=CC=CC=C1)OC1=C(C=C(C(=C1)[N+](=O)[O-])CC)OC Chemical compound C(C1=CC=CC=C1)OC1=C(C=C(C(=C1)[N+](=O)[O-])CC)OC FRCHGJBAEMHJOF-UHFFFAOYSA-N 0.000 description 2
- XOSMODLVKQELGP-UHFFFAOYSA-N C(C1=CC=CC=C1)OC1=CC(=C(C=C1OC)C(CO)C)[N+](=O)[O-] Chemical compound C(C1=CC=CC=C1)OC1=CC(=C(C=C1OC)C(CO)C)[N+](=O)[O-] XOSMODLVKQELGP-UHFFFAOYSA-N 0.000 description 2
- CCIIJBRRPBBRDO-UHFFFAOYSA-N COC=1C(=CC(=C(C1)C(CO)C)[N+](=O)[O-])OCC#C Chemical compound COC=1C(=CC(=C(C1)C(CO)C)[N+](=O)[O-])OCC#C CCIIJBRRPBBRDO-UHFFFAOYSA-N 0.000 description 2
- 241000252506 Characiformes Species 0.000 description 2
- 229910021589 Copper(I) bromide Inorganic materials 0.000 description 2
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-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
- PEEHTFAAVSWFBL-UHFFFAOYSA-N Maleimide Chemical compound O=C1NC(=O)C=C1 PEEHTFAAVSWFBL-UHFFFAOYSA-N 0.000 description 2
- RZAMEAJUHVKTSH-UHFFFAOYSA-N OC1=CC(=C(C=C1OC)C(CO)C)[N+](=O)[O-] Chemical compound OC1=CC(=C(C=C1OC)C(CO)C)[N+](=O)[O-] RZAMEAJUHVKTSH-UHFFFAOYSA-N 0.000 description 2
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- PVDVPOZEJCXUAM-UHFFFAOYSA-N acetonitrile;n,n-diethylethanamine Chemical compound CC#N.CCN(CC)CC PVDVPOZEJCXUAM-UHFFFAOYSA-N 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- AGEZXYOZHKGVCM-UHFFFAOYSA-N benzyl bromide Chemical compound BrCC1=CC=CC=C1 AGEZXYOZHKGVCM-UHFFFAOYSA-N 0.000 description 2
- 125000001246 bromo group Chemical group Br* 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 125000001309 chloro group Chemical group Cl* 0.000 description 2
- NKNDPYCGAZPOFS-UHFFFAOYSA-M copper(i) bromide Chemical compound Br[Cu] NKNDPYCGAZPOFS-UHFFFAOYSA-M 0.000 description 2
- KPUPVWORBVCEOT-UHFFFAOYSA-N copper;triphenylphosphane Chemical compound [Cu].C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 KPUPVWORBVCEOT-UHFFFAOYSA-N 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 125000003827 glycol group Chemical group 0.000 description 2
- VPFMEXRVUOPYRG-UHFFFAOYSA-N hex-5-ynoic acid Chemical compound OC(=O)CCCC#C VPFMEXRVUOPYRG-UHFFFAOYSA-N 0.000 description 2
- 239000000693 micelle Substances 0.000 description 2
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 2
- 125000006502 nitrobenzyl group Chemical group 0.000 description 2
- KJIFKLIQANRMOU-UHFFFAOYSA-N oxidanium;4-methylbenzenesulfonate Chemical compound O.CC1=CC=C(S(O)(=O)=O)C=C1 KJIFKLIQANRMOU-UHFFFAOYSA-N 0.000 description 2
- 238000001782 photodegradation Methods 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
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- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 2
- YUYCVXFAYWRXLS-UHFFFAOYSA-N trimethoxysilane Chemical compound CO[SiH](OC)OC YUYCVXFAYWRXLS-UHFFFAOYSA-N 0.000 description 2
- SPEOJCYNZBPAAY-UHFFFAOYSA-N (6-bromo-7-hydroxy-2-oxochromen-4-yl)methyl hex-5-ynoate Chemical compound O1C(=O)C=C(COC(=O)CCCC#C)C2=C1C=C(O)C(Br)=C2 SPEOJCYNZBPAAY-UHFFFAOYSA-N 0.000 description 1
- GYAWTXGPAGMMFQ-UHFFFAOYSA-N 11-azido-1,1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8-heptadecafluoroundecane Chemical compound FC(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)CCCN=[N+]=[N-] GYAWTXGPAGMMFQ-UHFFFAOYSA-N 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- QPSDFLMKLTXKDA-UHFFFAOYSA-N 3-azidopropyl(trimethoxy)silane Chemical compound CO[Si](OC)(OC)CCCN=[N+]=[N-] QPSDFLMKLTXKDA-UHFFFAOYSA-N 0.000 description 1
- PFEFOHMLVFUELJ-UHFFFAOYSA-N 3-sulfanylideneisoindol-1-one Chemical compound C1=CC=C2C(=O)NC(=S)C2=C1 PFEFOHMLVFUELJ-UHFFFAOYSA-N 0.000 description 1
- CVTWASMMVSOPEW-UHFFFAOYSA-N 4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,11-heptadecafluoroundecan-1-amine Chemical compound NCCCC(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)F CVTWASMMVSOPEW-UHFFFAOYSA-N 0.000 description 1
- MPCCNXGZCOXPMG-UHFFFAOYSA-N 4-bromobenzene-1,3-diol Chemical compound OC1=CC=C(Br)C(O)=C1 MPCCNXGZCOXPMG-UHFFFAOYSA-N 0.000 description 1
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- ANJBPMLSQSNNFX-UHFFFAOYSA-N C(CCC=C)(=O)OCC1=CC(OC2=CC(=C(C=C12)Br)O)=O Chemical compound C(CCC=C)(=O)OCC1=CC(OC2=CC(=C(C=C12)Br)O)=O ANJBPMLSQSNNFX-UHFFFAOYSA-N 0.000 description 1
- GZUKWZUHIBYWIC-UHFFFAOYSA-N C(CCC=C)(=O)OCC1=CC(OC2=CC(=C(C=C12)Br)OC(=O)OC(C)(C)C)=O Chemical compound C(CCC=C)(=O)OCC1=CC(OC2=CC(=C(C=C12)Br)OC(=O)OC(C)(C)C)=O GZUKWZUHIBYWIC-UHFFFAOYSA-N 0.000 description 1
- 101100493713 Caenorhabditis elegans bath-45 gene Proteins 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- HAXFWIACAGNFHA-UHFFFAOYSA-N aldrithiol Chemical compound C=1C=CC=NC=1SSC1=CC=CC=N1 HAXFWIACAGNFHA-UHFFFAOYSA-N 0.000 description 1
- 125000005370 alkoxysilyl group Chemical group 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 229920000469 amphiphilic block copolymer Polymers 0.000 description 1
- NDKBVBUGCNGSJJ-UHFFFAOYSA-M benzyltrimethylammonium hydroxide Chemical compound [OH-].C[N+](C)(C)CC1=CC=CC=C1 NDKBVBUGCNGSJJ-UHFFFAOYSA-M 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- PFYXSUNOLOJMDX-UHFFFAOYSA-N bis(2,5-dioxopyrrolidin-1-yl) carbonate Chemical compound O=C1CCC(=O)N1OC(=O)ON1C(=O)CCC1=O PFYXSUNOLOJMDX-UHFFFAOYSA-N 0.000 description 1
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- OHLRLMWUFVDREV-UHFFFAOYSA-N ethyl 4-chloro-3-oxobutanoate Chemical compound CCOC(=O)CC(=O)CCl OHLRLMWUFVDREV-UHFFFAOYSA-N 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
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- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
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- HVAMZGADVCBITI-UHFFFAOYSA-N pent-4-enoic acid Chemical compound OC(=O)CCC=C HVAMZGADVCBITI-UHFFFAOYSA-N 0.000 description 1
- NMHMNPHRMNGLLB-UHFFFAOYSA-N phloretic acid Chemical group OC(=O)CCC1=CC=C(O)C=C1 NMHMNPHRMNGLLB-UHFFFAOYSA-N 0.000 description 1
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Landscapes
- Pyrane Compounds (AREA)
- Pyrrole Compounds (AREA)
- Low-Molecular Organic Synthesis Reactions Using Catalysts (AREA)
Description
本発明は、両末端に異なる反応性基を備えるカップリング剤であって、二光子励起でも切断可能な光分解性カップリング剤に関する。 The present invention relates to a coupling agent having different reactive groups at both ends, which is capable of being cleaved even by two-photon excitation.
自己組織化単分子膜(self-assembled monolayer,SAM)を形成するシランカップリング剤(非特許文献1参照)やチオール系カップリング剤(非特許文献2参照)は、無機材料表面に有機物を固定化して表面を機能化するための表面修飾剤として有用である。片末端に有機官能基、もう一端に表面との結合基を有する構造をしており、シランカップリング剤ではシリル基がシリコンやガラスなどの表面にあるヒドロキシ基と反応し、またチオール系カップリング剤ではチオール基がスルフィド結合を介して金などの表面に結合する。これらカップリング剤に光分解性基を導入したものが感光性表面修飾剤である。感光性表面修飾剤を用いれば、基板表面の化学的および物理的性質(官能基の導入やぬれ性の変換など)を光により時空間制御することが可能となり、フォトマスクを介した光照射によりSAMにパターンを形成することができることから、ケミカル・バイオセンシングやエレクトロニクスなど多様な用途への応用が期待される。
また、ポリマー間を光分解性リンカーで連結し、ブロック間を光で容易に分解することを可能にしたものが光分解性ブロックコポリマーである。特に、親水性ブロックと疎水性ブロックから構成される両親媒性ブロックコポリマーは、例えば、水溶液中では自己集合して、外殻に親水性ブロックが配向して疎水性ブロックが内部に存在するように分子が集合したミセル構造や二重膜となったポリマーソーム構造を形成することが知られている。ミセルやポリマーソームは、内部に薬剤を保持させることができるため、薬剤などのキャリアーとしてドラッグデリバリーシステム(DDS)に有用である。DDSでは、薬剤の運搬だけではなく、目的地に到達した後には薬剤を放出することが重要となる。光で崩壊するミセルやポリマーソームであれば、任意の時に任意の場所で、容易に内包物を放出することが可能である。さらに、照射する光量を調節することで、内容物を徐放することも可能となる。
Silane coupling agents (see Non-Patent Document 1) and thiol-based coupling agents (see Non-Patent Document 2) that form self-assembled monolayers (SAMs) fix organic substances on the surface of inorganic materials. It is useful as a surface modifier for functionalizing the surface. It has a structure that has an organic functional group at one end and a bonding group to the surface at the other end. In the silane coupling agent, the silyl group reacts with the hydroxy group on the surface of silicon or glass, and thiol coupling. In the agent, the thiol group is bonded to the surface of gold or the like through a sulfide bond. A photosensitive surface modifier is obtained by introducing a photodegradable group into these coupling agents. By using a photosensitive surface modifier, it becomes possible to control the chemical and physical properties of the substrate surface (introduction of functional groups, conversion of wettability, etc.) by light and time, and by light irradiation through a photomask. Since patterns can be formed on SAM, it is expected to be applied to various uses such as chemical biosensing and electronics.
A photodegradable block copolymer is a polymer in which the polymers are connected by a photodegradable linker and the blocks can be easily decomposed by light. In particular, an amphiphilic block copolymer composed of a hydrophilic block and a hydrophobic block is self-assembled in an aqueous solution, for example, so that the hydrophilic block is oriented in the outer shell and the hydrophobic block exists inside. It is known to form a micellar structure in which molecules are assembled and a polymersome structure in a double membrane. Since micelles and polymersomes can hold drugs inside, they are useful as drug carriers in drug delivery systems (DDS). In DDS, it is important not only to transport the drug but also to release the drug after reaching the destination. In the case of micelles or polymersomes that are disintegrated by light, inclusions can be easily released at any time and at any place. Further, the contents can be released gradually by adjusting the amount of light to be irradiated.
光分解性基は有機合成のために光で脱保護できる保護基として開発された(非特許文献3参照)。光分解性保護基の代表的なものとしては2−ニトロベンジル基が知られており、固相合成のリンカーとしての使用(非特許文献4参照)、また最近注目されているのがケージド化合物としての利用である(非特許文献5参照)。ケージド化合物とは、生理活性分子を光分解性基で化学修飾することで一時的に活性を失わせ、光で脱保護することで活性を覚醒させることのできる化合物の総称である。
有機合成の保護基としてはさほど注目されなかった光分解性基であるが、光による局所的制御に適することからその利用は近年になって再認識されている。
The photodegradable group was developed as a protecting group that can be deprotected with light for organic synthesis (see Non-Patent Document 3). A typical photodegradable protecting group is a 2-nitrobenzyl group, which is used as a linker for solid-phase synthesis (see Non-Patent Document 4), and recently a caged compound is attracting attention. (See Non-Patent Document 5). A caged compound is a generic term for compounds that can temporarily lose activity by chemically modifying a bioactive molecule with a photodegradable group and awaken the activity by deprotection with light.
Although it is a photodegradable group that has not received much attention as a protective group for organic synthesis, its use has been recognized again in recent years because it is suitable for local control by light.
また、従来は紫外線を用いた一光子励起による光分解のみであったが、近赤外線を用いた二光子励起による光分解が可能な光分解性保護基も注目されている(非特許文献6〜8)。二光子励起では、エネルギーの低い近赤外パルスレーザーを光源とし、光源を集光することによって光子密度が非常に高くなる焦点領域のみで二つの光子を吸収することにより励起され、光分解性基が切断される。そのため、非常に高い三次元分解能を実現できる。すなわち、焦点の位置を制御することにより、三次元の微細パターンを精密に作製することも可能となる。一方、従来の一光子励起では、エネルギーの高い紫外光などを光源とするため、光源の焦点以外の領域でも光分解が起こるために分解能は低い。 Conventionally, only photodecomposition by one-photon excitation using ultraviolet rays has been performed, but photodegradable protecting groups capable of photolysis by two-photon excitation using near infrared rays have been attracting attention (Non-Patent Documents 6 to 6). 8). In two-photon excitation, a low-energy near-infrared pulsed laser is used as a light source, and it is excited by absorbing two photons only in the focal region where the photon density becomes very high by condensing the light source. Is disconnected. Therefore, very high three-dimensional resolution can be realized. That is, by controlling the position of the focal point, a three-dimensional fine pattern can be accurately produced. On the other hand, in the conventional one-photon excitation, high-energy ultraviolet light or the like is used as a light source, so that photolysis occurs even in a region other than the focal point of the light source, so the resolution is low.
光分解性保護基として用いられる2−ニトロベンジル基は、光分解でニトロソ体を生じるため、ケージ解除後の副生成物に毒性があり、また、光分解でアルデヒドあるいはケトンを生じるため、副反応の恐れがある(非特許文献9)。さらには、二光子励起による光分解効率が悪いため、一光子励起による光分解では細胞等の生体材料に対してダメージが懸念されると共に2次元的なパターニングのみしか可能でない、等の問題点が指摘されている。 The 2-nitrobenzyl group used as a photodegradable protecting group generates a nitroso form by photolysis, and therefore is toxic to by-products after releasing the cage, and also generates an aldehyde or a ketone by photolysis. (Non-patent document 9). Furthermore, since the photolysis efficiency by two-photon excitation is poor, there is a problem that the photolysis by one-photon excitation may cause damage to biological materials such as cells and only two-dimensional patterning is possible. It has been pointed out.
また、本出願人は、先に、同一分子内に異なる反応性基を備えるヘテロ二価性のカップリング剤であって、加水分解性シリル基、アミン反応性基、及びチオール反応性基からなる群から選択される2つの異なる反応性基を備え、この両反応性基の間に光分解性基を備えた光分解性カップリング剤(具体的には、一方の末端に加水分解性シリル基を有し、他方の末端にアミン反応性基又はチオール反応性基を有し、これら両末端基の間に光分解性基を含む光分解性カップリング剤、又は、一方の末端にアミン反応性基を有し、他方の末端にチオール反応性基を有し、これら両末端基の間に光分解性基を含む光分解性カップリング剤)や(特開2007−186472号公報)、同一分子内に、アジド反応性基(アルキン)又はアルキン反応性基(アジド)と、アミン反応性基又はチオール反応性基とを同一分子内にスペーサを介して備えるヘテロ二価性架橋剤であって、一方の末端にクリックケミストリーによりアジドと反応するアルキン、又は、アルキンと反応するアジドを含み、他末端にアミン反応性基又はチオール反応性基を含み、これら両反応性基の間に光分解性基を設けた光分解性へテロ二価性架橋剤(特開2010−260831号公報)について提案している。
これらの光分解性カップリング剤や光分解性ヘテロ二価性架橋剤にあっても、従来の光分解で用いていた紫外光の2倍の波長の可視光(波長700〜800nm)を光源とする二光子励起による光分解が可能となれば、細胞などの生体材料に対してダメージが少なくなり、また、2次元でなく3次元での材料のパターニングが可能になるなど、利用用途が大きく広がることとなる。
Further, the applicant of the present invention is a heterobivalent coupling agent having different reactive groups in the same molecule, and comprises a hydrolyzable silyl group, an amine reactive group, and a thiol reactive group. A photodegradable coupling agent comprising two different reactive groups selected from the group, and a photodegradable group between both reactive groups (specifically, a hydrolyzable silyl group at one end) A photodegradable coupling agent having an amine-reactive group or a thiol-reactive group at the other end, and a photodegradable group between these two end groups, or an amine-reactive at one end A photodegradable coupling agent having a group, a thiol-reactive group at the other end, and a photodegradable group between these both end groups) (Japanese Patent Laid-Open No. 2007-186472), the same molecule Inside, azide reactive group (alkyne) or alkyne reactive group An azide) and an amine-reactive group or thiol-reactive group in the same molecule through a spacer, an alkyne that reacts with azide at one end by click chemistry, or an alkyne A photodegradable heterobivalent cross-linking agent comprising an azide that reacts with an amino group, an amine-reactive group or a thiol-reactive group at the other end, and a photodegradable group provided between these reactive groups 2010-260831).
Even in these photodegradable coupling agents and photodegradable heterobivalent crosslinking agents, visible light (wavelength 700 to 800 nm) having a wavelength twice that of ultraviolet light used in conventional photolysis is used as a light source. If photolysis by two-photon excitation is possible, damage to biological materials such as cells will be reduced, and patterning of materials in three dimensions instead of two dimensions will be possible, and the usage will be greatly expanded. It will be.
本発明は係る事情に鑑みてなされたものであり、光分解による分解生成物の毒性が低く、また、光分解によって生じる生成物による副反応の恐れがなく、さらには、近赤外線による二光子励起による光分解を可能として、光分解時に生体材料へのダメージを少なくすると共に一光子励起に比べて高い3次元の空間分解能で局所的に照射することを可能とし、さらには、従来の光分解性カップリング剤や光分解性ヘテロ二価性架橋剤においても二光子励起による光分解を可能として利用用途を拡大することが可能な新規な光分解性カップリング剤を提供することを主たる課題としている。 The present invention has been made in view of the circumstances, and the toxicity of the decomposition product by photolysis is low, there is no fear of side reaction by the product caused by photolysis, and further, two-photon excitation by near infrared rays Can be photodegraded, reducing damage to biomaterials during photodecomposition, enabling local irradiation with higher three-dimensional spatial resolution than one-photon excitation, and conventional photodegradability The main task is to provide a novel photodegradable coupling agent that can be used for photolysis by two-photon excitation and can be used for a coupling agent and a photodegradable heterobivalent crosslinking agent. .
上記課題を達成するために、本発明に係る光分解性カップリング剤は、両末端に反応性基を有し、その間にクマリニルメチル型光分解性基を有することを特徴としている。 In order to achieve the above object, the photodegradable coupling agent according to the present invention is characterized by having a reactive group at both ends and a coumarinylmethyl type photodegradable group between them.
具体的には、下記一般式(1)
以上述べたクマリニルメチル型光分解性基を備えた光分解性カップリング剤によれば、以下の諸効果が得られる。
(1)光分解でニトロソ体を生じないため、光照射後の生成物の毒性が低い。
(2)光分解でアルデヒドまたはケトンを生じないため、副反応の恐れがない。
(3)一光子励起(紫外線)に加えて、二光子励起(近赤外線)による光分解も可能であるため、長波長(1/2のエネルギー)の光で切断が可能であり、細胞等の生体材料へのダメージが少なく、また、1光子励起に比べて高い3次元の空間分解能で局所的に照射することが可能となり、3次元での材料のパターニングが可能である。
(4)従来の光分解性カップリング剤や光分解性ヘテロ二価性架橋剤においても二光子励起による光分解を可能として利用用途を拡大することが可能となる。
According to the photodegradable coupling agent provided with the coumarinylmethyl type photodegradable group described above, the following effects are obtained.
(1) Since the nitroso form is not generated by photolysis, the toxicity of the product after light irradiation is low.
(2) Since no aldehyde or ketone is produced by photolysis, there is no risk of side reactions.
(3) In addition to one-photon excitation (ultraviolet), photodecomposition by two-photon excitation (near infrared) is also possible, so it can be cleaved with light of long wavelength (1/2 energy), There is little damage to the biomaterial, and it is possible to irradiate locally with a higher three-dimensional spatial resolution than one-photon excitation, and patterning of the material in three dimensions is possible.
(4) Conventional photodegradable coupling agents and photodegradable heterobivalent cross-linking agents can also be used in a wider range of applications by enabling photodecomposition by two-photon excitation.
以下、この発明の実施形態を説明する。
本発明の光分解性カップリング剤は、両末端に異なる反応性基を備え、その間に、ニトロベンジル型光分解性基に代えて、ニトロフェネチル型光分解性基(参考例)またはクマリニルメチル型光分解性基を備えたものである。
Embodiments of the present invention will be described below.
The photodegradable coupling agent of the present invention has different reactive groups at both ends, and in the meantime, instead of a nitrobenzyl type photodegradable group, a nitrophenethyl type photodegradable group (reference example) or coumarinylmethyl Type photodegradable group.
ニトロフェネチル型光分解性基を両末端の反応性基の間に含む光分解性カップリング剤としては、下記の一般式(1)で示される2−(5−メトキシ−2−ニトロ−4−プロパ−2−インイルオキシフェニル)プロピル N−スクシンイミジルカーボネートが考えられる。 As a photodegradable coupling agent containing a nitrophenethyl-type photodegradable group between the reactive groups at both ends, 2- (5-methoxy-2-nitro-4-) represented by the following general formula (1) is used. Propa-2-ynyloxyphenyl) propyl N-succinimidyl carbonate is conceivable.
また、クマリニルメチル型光分解性基を両末端の反応性基の間に含む光分解性カップリング剤としては、下記の一般式(2)で示される光分解性カップリング剤や、一般式(3)で示される光分解性カップリング剤が考えられる。
このような光分解性カップリング剤を用いることにより、従来のニトロベンジル型光分解性基と比較すると、光分解でニトロソ体を生じないので毒性が低く、また、光分解でアルデヒド又はケトンを生じないので、副反応がない。
また、一光子励起(紫外線)での光分解に加えて、二光子励起(近赤外線)による光分解も可能である。このため、2倍の長波長の光、即ち、約半分のエネルギーの光で切断が可能であり、また、2次元ではなく、3次元での分解能が得られる(3次元での材料のパターニングが可能となる)。
By using such a photodegradable coupling agent, compared with the conventional nitrobenzyl type photodegradable group, the photodecomposition does not produce a nitroso form, so the toxicity is low, and photodegradation produces an aldehyde or a ketone. Because there is no side reaction.
In addition to photodecomposition by one-photon excitation (ultraviolet light), photodecomposition by two-photon excitation (near infrared) is also possible. For this reason, it is possible to cut with twice the wavelength of light, that is, about half of the energy, and it is possible to obtain a resolution in three dimensions instead of two dimensions (material patterning in three dimensions is possible). Possible).
以下において、上述した光分解性をより具体的に説明するが、本発明は、これに限定されるものではない。尚、以下の例において、水とはイオン交換水を指す。 In the following, the above-described photodegradability will be described more specifically, but the present invention is not limited to this. In the following examples, water refers to ion exchange water.
ニトロフェネチル型光分解性カップリング剤の例として、2−(5−メトキシ−2−ニトロ−4−プロパ−2−インイルオキシフェニル)プロピル N−スクシンイミジルカーボネートを示す。 As an example of a nitrophenethyl type photodegradable coupling agent, 2- (5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl) propyl N-succinimidyl carbonate is shown.
このニトロフェネチル型光分解性カップリング剤を図1に示す工程により合成した。
先ず、500 mLナスフラスコに4-ethylguaiacal 5.34 g (35.1 mmol, 1.0 eq)をacetone 200 mLに溶かし、K2CO3 56.7 g (410 mmol, 2.5 eq)を加えて1時間攪拌した。その後、benzyl bromide 39 mL (328 mmol, 2.0 eq)を加えて14時間還流した(オイルバス温度70 ℃)。反応液を濃縮後、H2O 200 mLを加え、chloroform (200 mL×3)で抽出し、anhydrous MgSO4で乾燥、ろ過、濃縮、真空乾燥した。減圧蒸留でbenzyl bromideを留去し、淡黄色オイル37.2 g (154 mmol)を得た(ステップ1)。
This nitrophenethyl type photodegradable coupling agent was synthesized by the process shown in FIG.
First, 5.34 g (35.1 mmol, 1.0 eq) of 4-ethylguaiacal was dissolved in 200 mL of acetone in a 500 mL eggplant flask, and 56.7 g (410 mmol, 2.5 eq) of K 2 CO 3 was added and stirred for 1 hour. Thereafter, 39 mL (328 mmol, 2.0 eq) of benzyl bromide was added and refluxed for 14 hours (oil bath temperature 70 ° C.). After the reaction solution was concentrated, 200 mL of H 2 O was added, extracted with chloroform (200 mL × 3), dried over anhydrous MgSO 4 , filtered, concentrated, and vacuum dried. The benzyl bromide was distilled off under reduced pressure to obtain 37.2 g (154 mmol) of pale yellow oil (Step 1).
上記合成で得られた1−ベンジルオキシ−4−エチル−2−メトキシベンゼンの同定結果を以下に示す。
The identification results of 1-benzyloxy-4-ethyl-2-methoxybenzene obtained by the above synthesis are shown below.
次に、100 mL二口ナスフラスコに窒素雰囲気下において、nitronium tetrafluoroborate 0.629 g (4.73 mmol, 1.1 eq)を入れ、dry-acetonitrile 20 mLで溶解した。フラスコに2-picoline 0.467 mL (0.440 g, 0.473 mmol, 1.2 eq)をdry-acetonitrile 20 mLで溶解し、-30 ℃下において、滴下ロートを用いて0.5時間かけて滴下し、さらに0.5 時間攪拌した。1-benzyloxy-4-ethyl-2-methoxybenzene 1.05 g (4.30 mmol, 1.0 eq)をdry-acetonitrile 10 mLで溶解し、滴下ロートを用いて1時間かけて滴下し、ゆっくりと室温に戻して12 時間攪拌した。濃縮後、sat. NaHCO3 aq. 100 mLを加え、ethyl acetate (100 mL×3)で抽出した。その後anhydrous MgSO4で乾燥、ろ過、濃縮、真空乾燥を行い、黄褐色固体1.30 gを得た。ethyl acetateで再結晶し、第一結晶0.654 g ( 2.28 mmol)、第二結晶0.222 g ( 0.777 mmol)を得た。また第三結晶をカラムクロマトグラフィー(hexane : ethyl acetate = 5 : 1 [ カラム径 3 cm シリカ長 15 cm ])で単離精製し、濃縮、真空乾燥を行い、黄色固体0.184 gを得た(ステップ2)。 Next, nitronium tetrafluoroborate 0.629 g (4.73 mmol, 1.1 eq) was placed in a 100 mL two-necked eggplant flask under a nitrogen atmosphere, and dissolved in 20 mL of dry-acetonitrile. 2-picoline 0.467 mL (0.440 g, 0.473 mmol, 1.2 eq) was dissolved in a flask with 20 mL of dry-acetonitrile, added dropwise with a dropping funnel over 0.5 hour at −30 ° C., and further stirred for 0.5 hour. . Dissolve 1.05 g (4.30 mmol, 1.0 eq) of 1-benzyloxy-4-ethyl-2-methoxybenzene in 10 mL of dry-acetonitrile, add dropwise over 1 hour using a dropping funnel, and slowly return to room temperature for 12 hours. Stir. After concentration, sat. NaHCO 3 aq. 100 mL was added, and the mixture was extracted with ethyl acetate (100 mL × 3). Thereafter, drying with anhydrous MgSO 4 , filtration, concentration, and vacuum drying were performed to obtain 1.30 g of a tan solid. Recrystallization from ethyl acetate gave 0.654 g (2.28 mmol) of the first crystal and 0.222 g (0.777 mmol) of the second crystal. The third crystal was isolated and purified by column chromatography (hexane: ethyl acetate = 5: 1 [column diameter 3 cm, silica length 15 cm]), concentrated and vacuum dried to obtain 0.184 g of a yellow solid (step 2).
上記合成で得られた1−ベンジルオキシ−4−エチル−2−メトキシ−5−ニトロベンゼンの同定結果を以下に示す。
The identification results of 1-benzyloxy-4-ethyl-2-methoxy-5-nitrobenzene obtained by the above synthesis are shown below.
次に、300 mL 二口ナスフラスコに窒素雰囲気下において、1-benzyloxy-4-ethyl-2-methoxy-5-nitrobenzene 7.21 g (25.0 mmol, 1.0 eq)、paraformaldehyde 3.42 g (104 mmol, 4.1 eq)、dry-dimethylslufoxide 40 mLを入れ、dry-dimethylslufoxide 5 mLに溶かしたTriton B 2.76 mL (2.93 g, 17.5 mmol, 0.7 eq)を滴下し、17時間攪拌した。その後、sat.NH4Cl aq. 100 mL加え、ethyl acetate (100 mL×3)で抽出し、有機層をsat.NaCl aq. (×3)で洗浄、anhydrous MgSO4で乾燥、ろ過、濃縮、真空乾燥を行い、黄色オイル7.95 g を得た。その後カラムクロマトグラフィー(hexane : ethyl acetate = 1 : 1 [ カラム径 6 cm シリカ長 15 cm ]) で単離精製し、濃縮、真空乾燥を行い、黄色オイル 6.19 g (19.5 mmol)を得た(ステップ3)。 Next, in a 300 mL two-necked eggplant flask under nitrogen atmosphere, 1-benzyloxy-4-ethyl-2-methoxy-5-nitrobenzene 7.21 g (25.0 mmol, 1.0 eq), paraformaldehyde 3.42 g (104 mmol, 4.1 eq) Then, 40 mL of dry-dimethylslufoxide was added, and 2.76 mL (2.93 g, 17.5 mmol, 0.7 eq) of Triton B dissolved in 5 mL of dry-dimethylslufoxide was added dropwise, followed by stirring for 17 hours. Then, sat.NH 4 Cl aq. 100 mL was added, extracted with ethyl acetate (100 mL × 3), the organic layer was washed with sat.NaCl aq. (× 3), dried over anhydrous MgSO 4 , filtered, concentrated, Vacuum drying was performed to obtain 7.95 g of a yellow oil. After that, it was isolated and purified by column chromatography (hexane: ethyl acetate = 1: 1 [column diameter 6 cm, silica length 15 cm]), concentrated and vacuum dried to obtain 6.19 g (19.5 mmol) of yellow oil (step 3).
上記合成で得られた2−(4−ベンジルオキシ−5−メトキシ−2−ニトロフェニル)プロパノールの同定結果を以下に示す。
The identification result of 2- (4-benzyloxy-5-methoxy-2-nitrophenyl) propanol obtained by the above synthesis is shown below.
次に、200 mLナスフラスコに、2-(4-benzyloxy-5-methoxy-2-nitrophenyl)propanol 6.19 g (19.5 mmol)、tetrahydrofrane 40 mLを加え、conc.HCl aq. 130 mLを滴下ロートで滴下し、室温で16時間攪拌した。反応溶液に、5% NaHCO3 aq. 250 mLを加え、ethyl acetate (250 mL×3)で抽出し、続いて有機層をsat. NaCl aq. (×3)で洗浄、anhydrous MgSO4 で乾燥、濃縮、真空乾燥を行い、緑色固体8.81 gを得た。ethyl acetateで再結晶を行い、第一結晶9.96 g (34.7 mmol)、第二結晶3.50 g (12.2 mmol)、また第二結晶のろ液をカラムクロマトグラフィー(hexane : ethyl acetate = 1 : 1 [ カラム径 5 cm シリカ長 20 cm ])で単離精製し、濃縮、真空乾燥を行い、黄色固体0.521 g を得た(ステップ4)。 Next, to a 200 mL eggplant flask, add 6.19 g (19.5 mmol) of 2- (4-benzyloxy-5-methoxy-2-nitrophenyl) propanol and 40 mL of tetrahydrofrane, and add conc.HCl aq. 130 mL with a dropping funnel. And stirred at room temperature for 16 hours. To the reaction solution, add 5% NaHCO 3 aq. 250 mL, extract with ethyl acetate (250 mL × 3), then wash the organic layer with sat. NaCl aq. (× 3), dry over anhydrous MgSO 4 Concentration and vacuum drying were performed to obtain 8.81 g of a green solid. Recrystallize with ethyl acetate, and column chromatography (hexane: ethyl acetate = 1: 1: column) of the first crystal 9.96 g (34.7 mmol), the second crystal 3.50 g (12.2 mmol), and the second crystal filtrate. It was isolated and purified with a diameter of 5 cm and a silica length of 20 cm]), concentrated and vacuum dried to obtain 0.521 g of a yellow solid (Step 4).
上記合成で得られた2−(4−ヒドロキシ−5−メトキシ−2−ニトロフェニル)プロパノールの同定結果を以下に示す。
The identification result of 2- (4-hydroxy-5-methoxy-2-nitrophenyl) propanol obtained by the above synthesis is shown below.
次に、100 mL二口ナスフラスコに窒素雰囲気下において、2-(4-hydroxy-5-methoxy-2-nitrophenyl)propanol 1.01 g (4.44 mmol, 1.0 eq)、dry-acetonitrile 20 mL、K2CO3を1.22 g (8.87 mmol, 2.0 eq)加え、室温で1時間攪拌した。その後、propargyl bromide 0.504 mL (0.792 g, 6.65 mmol, 1.5 eq)を加え、14.5時間還流した(オイルバス温度90℃)。その後、反応溶液にsat. NH4Cl aq. 30 mL, 2N HCl aq. 20 mLを加え、ethyl acetate (50 mL×3)で抽出し、続いて有機層をsat. NaCl aq. (×3)で洗浄、anhydrous MgSO4で乾燥、濃縮、真空乾燥を行い、黄色固体1.20 gを得た。カラムクロマトグラフィー(hexane : ethyl acetate = 1 : 1 [カラム径 3 cm シリカ長 15 cm])を行い、黄色固体1.17 g (4.40 mmol)を得た(ステップ5)。 Next, in a 100 mL two-necked eggplant flask under nitrogen atmosphere, 2- (4-hydroxy-5-methoxy-2-nitrophenyl) propanol 1.01 g (4.44 mmol, 1.0 eq), dry-acetonitrile 20 mL, K 2 CO 1.22 g (8.87 mmol, 2.0 eq) of 3 was added, and the mixture was stirred at room temperature for 1 hour. Then, propargyl bromide 0.504 mL (0.792 g, 6.65 mmol, 1.5 eq) was added and refluxed for 14.5 hours (oil bath temperature 90 ° C.). Then, sat. NH 4 Cl aq. 30 mL, 2N HCl aq. 20 mL was added to the reaction solution, extracted with ethyl acetate (50 mL × 3), and then the organic layer was washed with sat. NaCl aq. (× 3) After washing with, dried over anhydrous MgSO 4 , concentrated and vacuum dried to obtain 1.20 g of a yellow solid. Column chromatography (hexane: ethyl acetate = 1: 1 [column diameter 3 cm, silica length 15 cm]) was performed to obtain 1.17 g (4.40 mmol) of a yellow solid (Step 5).
上記合成で得られた2−(5−メトキシ−2−ニトロ−4−プロパ−2−インイルオキシフェニル)プロパノールの同定結果を以下に示す。
The identification result of 2- (5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl) propanol obtained by the above synthesis is shown below.
次に、20 mL 二口ナスフラスコに窒素雰囲気下において、2-(5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl)propanol 126 mg (0.475 mmol, 1.0eq)、dry-acetonitrile 10 mL, N,N-disuccinimidyl carbonate 370 mg (1.43 mmol, 3.0 eq)を入れ、triethylamine 0.199 mL (1.43 mmol, 3.0 eq)を入れ、室温で22.5時間攪拌した。反応溶液に、sat. NH4Cl aq. 50 mLを加え、chloroform (50 mL×3)で抽出し、続いて有機層をsat. NaHCO3 aq. (×3)で洗浄、anhydrous MgSO4 で乾燥、濃縮、真空乾燥を行い、黄色オイル 346 mgを得た。カラムクロマトグラフィー(dichloromethane : ethyl acetate = 10 : 1 [カラム径 2 cm シリカ長 15 cm])を行い、淡黄色固体170 mg (0.418 mmol)を得た(ステップ6)。 Next, in a 20 mL double-necked eggplant flask under a nitrogen atmosphere, 2- (5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl) propanol 126 mg (0.475 mmol, 1.0eq), dry-acetonitrile 10 mL , N, N-disuccinimidyl carbonate 370 mg (1.43 mmol, 3.0 eq) was added, triethylamine 0.199 mL (1.43 mmol, 3.0 eq) was added, and the mixture was stirred at room temperature for 22.5 hours. To the reaction solution, add sat. NH 4 Cl aq. 50 mL, extract with chloroform (50 mL × 3), wash the organic layer with sat. NaHCO 3 aq. (× 3), and dry with anhydrous MgSO 4 Then, concentration and vacuum drying were performed to obtain 346 mg of a yellow oil. Column chromatography (dichloromethane: ethyl acetate = 10: 1 [column diameter 2 cm, silica length 15 cm]) was performed to obtain 170 mg (0.418 mmol) of a pale yellow solid (step 6).
上記合成で得られた2−(5−メトキシ−2−ニトロ−4−プロパ−2−インイルオキシフェニル)プロピル N−スクシンイミジルカーボネートの同定結果を以下に示す。
The identification results of 2- (5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl) propyl N-succinimidyl carbonate obtained by the above synthesis are shown below.
以上の工程で得られた光分解性カップリング剤としての2−(5−メトキシ−2−ニトロ−4−プロパ−2−インイルオキシフェニル)プロピル N−スクシンイミジルカーボネートを用いて表面修飾を行う場合の例を図2に示す。図2(a)は、アミノ基を含むシランカップリング剤を用いて基板表面上に単分子膜を形成し、このアミノ基に上述した光分解性カップリング剤の一方の反応基を結合させたものであり、図2(b)は、アルキン反応性基(アジド基)を含むシランカップリング剤を用いて基板表面上に単分子膜を形成し、このアジド基に上述した光分解性カップリング剤の他方の反応基を結合させるものである。さらに、光分解性カップリング剤を修飾した各基板において表面上に残るもう一方の反応性基を利用して、図2(a)においてはアルキニル基に対してアジド化合物を修飾することができ、図2(b)においてはスクシンイミジル基に対してアミン化合物を修飾することができる。 Surface modification using 2- (5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl) propyl N-succinimidyl carbonate as a photodegradable coupling agent obtained in the above steps FIG. 2 shows an example of performing the above. In FIG. 2A, a monomolecular film is formed on the substrate surface using a silane coupling agent containing an amino group, and one reactive group of the above-described photodegradable coupling agent is bonded to this amino group. FIG. 2 (b) shows a monomolecular film formed on the substrate surface using a silane coupling agent containing an alkyne-reactive group (azide group), and the photodegradable coupling described above is applied to this azide group. The other reactive group of the agent is bonded. Furthermore, using the other reactive group remaining on the surface of each substrate modified with the photodegradable coupling agent, the azide compound can be modified with respect to the alkynyl group in FIG. In FIG. 2B, the amine compound can be modified with respect to the succinimidyl group.
先ず、図2(a)の場合の実施例を示す。
先ず、シリコンウェハー基板をH2O、methanolで超音波洗浄し、窒素気流により乾燥させた後、UVオゾンクリーナーを用いて前処理をした。次いで、50 mL太口ナスフラスコに窒素雰囲気下においてdry-toluene 10 mL、3-aminopropyltrimethoxysilane 1.76 μL (1.79 mg, 10.0 μmol)を入れ、1 mM toluene溶液を調製した。その溶液に前処理済みのシリコンウェハー基板を入れ、100 ℃で4時間浸漬させた。その後methanolで洗い、methanolで超音波洗浄(10 min)を行った。次いでchloroformで洗い、chloroformで超音波洗浄(10 min)を行ない、表面を窒素気流で乾燥させた。この基板表面の水の接触角は46°であり、基板上にアミノ基が導入されたと考えられる。
さらに、10 mL二口試験管に窒素気流下においてdry-dimethylsulfoxde 6 mL入れ、2-(5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl)propyl N-succinimidyl carbonate 60 mg (0.148 mmol)、triethylamine 60 μL (8.17 mmol)加え、1%DMSO溶液を調製した。その溶液に前記のアミノ化基板を入れ、室温で23時間浸漬した。その後methanolで洗い、methanolで超音波洗浄(10 min)を行った。次いでchloroformで洗い、chloroformで超音波洗浄(10 min)を行ない、表面を窒素気流で乾燥させた。この基板表面の水の接触角は71°であり、水の接触角が上昇したことから疎水性になったことを示し、基板上に光分解性リンカーが導入されたことが考えられる。
First, an embodiment in the case of FIG.
First, the silicon wafer substrate was ultrasonically cleaned with H 2 O and methanol, dried with a nitrogen stream, and then pretreated with a UV ozone cleaner. Next, 10 mL of dry-toluene and 1.76 μL (1.79 mg, 10.0 μmol) of 3-aminopropyltrimethoxysilane were placed in a 50 mL large-mouth eggplant flask under a nitrogen atmosphere to prepare a 1 mM toluene solution. A pretreated silicon wafer substrate was placed in the solution and immersed at 100 ° C. for 4 hours. Then, it was washed with methanol and ultrasonically washed with methanol (10 min). Next, it was washed with chloroform, ultrasonically washed with chloroform (10 min), and the surface was dried with a nitrogen stream. The contact angle of water on the substrate surface is 46 °, and it is considered that an amino group was introduced onto the substrate.
Furthermore, put 6 mL of dry-dimethylsulfoxde in a 10 mL two-necked test tube under a nitrogen stream, and 2- (5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl) propyl N-succinimidyl carbonate 60 mg (0.148 mmol) Triethylamine 60 μL (8.17 mmol) was added to prepare a 1% DMSO solution. The aminated substrate was placed in the solution and immersed for 23 hours at room temperature. Then, it was washed with methanol and ultrasonically washed with methanol (10 min). Next, it was washed with chloroform, ultrasonically washed with chloroform (10 min), and the surface was dried with a nitrogen stream. The contact angle of water on the surface of the substrate was 71 °, indicating that the contact angle of water was increased, indicating that the substrate became hydrophobic, and it is considered that a photodegradable linker was introduced on the substrate.
その後、2−(5−メトキシ−2−ニトロ−4−プロパ−2−インイルオキシフェニル)プロピル N−スクシンイミジルカーボネートを表面に導入した修飾基板の光分解を調べるために、超高圧水銀灯で、320 nm以下の波長の光を遮断する硫酸銅フィルターを通して一定時間ごとに光照射し(波長 > 320 nm、光量 50 mW/ cm2)、水の接触角測定を行った。
光分解は次式のようになされ、図3に示す水の接触角測定より、時間と共に光分解が進行していることが確認された。
Then, in order to investigate the photolysis of the modified substrate into which 2- (5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl) propyl N-succinimidyl carbonate was introduced on the surface, an ultrahigh pressure mercury lamp Then, the contact angle of water was measured by irradiating light at regular intervals through a copper sulfate filter that blocks light with a wavelength of 320 nm or less (wavelength> 320 nm, light amount 50 mW / cm 2 ).
The photolysis was performed as follows, and it was confirmed from the measurement of the contact angle of water shown in FIG.
さらに、上述の工程で得られた光分解性カップリング剤を修飾した基板を用いて、他方の反応基であるアルキニル基に対するアジド化合物の修飾も行った。すなわち、50 mL 太口ナスフラスコに窒素雰囲気下において、4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,11-heptadecafluoroundecyl azide 12.5 mg(24.8 μmol)、dry-tetrahydrofuran 10 mLを入れ、約2 mM混合溶液を調製した。混合溶液に bromotris(triphenylphosphine)copper 14.3 mg (15.2 μmol), N,N-diisopropylethylamine 50 μL(36.5 mg , 0.282 mmol)を加え、リンカーを修飾した基板を入れ、60 ℃で25時間浸漬した。その後methanolで洗い、methanolで超音波洗浄(10 min)を行った。次いでchloroformで洗い、chloroformで超音波洗浄(10 min)を行ない、表面を窒素気流で乾燥させた。この基板表面の水の接触角は97°であり、大きく上昇したことから疎水性であるフッ化アルキル鎖が基板上に導入されたことを示す。また、図5に示すXPS測定からもフッ化アルキル鎖が導入されたことを確認した。
その後、得られた修飾基板の光分解を調べるために、超高圧水銀灯で、320 nm以下の波長の光を遮断する硫酸銅フィルターを通して一定時間ごとに光照射し(波長> 320 nm、光量 50 mW/ cm2)、水の接触角測定を行った。
光分解は次式のようになされ、図4に示す水の接触角測定より、時間と共に光分解が進行していることが確認された。また、図5に示すXPS測定において、光照射後にフッ化アルキル鎖及びニトロ基由来のピークが消失していることからも光分解を確認した。
The photolysis was performed as follows, and it was confirmed from the measurement of the contact angle of water shown in FIG. 4 that the photolysis progressed with time. Further, in the XPS measurement shown in FIG. 5, photolysis was also confirmed from the disappearance of the peaks derived from alkyl fluoride chains and nitro groups after light irradiation.
また、光分解性カップリング剤を修飾した基板に対して、上述とは異なるアジド化合物の修飾も行った。すなわち、50 mL太口ナスフラスコに窒素雰囲気下において、PEG350-azide 11.8 mg(31.5 μmol)、dry-tetrahydrofuran 10 mLを入れ、約3 mM混合溶液を調製した。混合溶液にbromotris(triphenylphosphine)copper 12.1 mg(13.0 μmol), N,N-diisopropylethylamine 60 μL(44.5 mg, 0.344 mmol)を加え、カップリング剤を修飾した基板を入れ、60 ℃で24時間浸漬した。その後methanolで洗い、methanolで超音波洗浄(10 min)を行った。次いでchloroformで洗い、chloroformで超音波洗浄(10 min)を行ない、表面を窒素気流で乾燥させた。この基板表面の水の接触角は48°であり、大きく減少したことから親水性であるポリエチレングリコール鎖が基板上に導入されたことを示す。
その後、得られた修飾基板の光分解を調べるために、超高圧水銀灯で、320 nm以下の波長の光を遮断する硫酸銅フィルターを通して一定時間ごとに光照射し(波長> 320 nm、光量50 mW/ cm2)、水の接触角測定を行った。
光分解は次式のようになされ、図6に示す水の接触角測定より、時間と共に光分解が進行していることが確認された。また、図7に示すXPS測定において、光照射後にニトロ基由来のピークが消失していることからも光分解を確認した。
The photolysis was performed according to the following equation, and it was confirmed from the measurement of the contact angle of water shown in FIG. Further, in the XPS measurement shown in FIG. 7, photolysis was also confirmed from the disappearance of the peak derived from the nitro group after light irradiation.
次に、図2(b)の場合の実施例を示す。
先ず、シリコンウェハー基板をH2O、methanolで超音波洗浄し、窒素気流により乾燥させた後、UVオゾンクリーナーを用いて前処理をした。次いで、50 ml太口ナスフラスコに(3-azidopropyl)trimethoxysilane 17.5 mg (85.2 μmol)入れ、tetrahydrofuran 4 mL、2N HCl 10 μL、cyclohexane 16 mL入れた。その溶液に前処理済みのシリコンウェハー基板を入れ、室温で18.5時間浸漬した。その後methanolで洗い、methanolで超音波洗浄(10 min)を行った。さらにchloroformで洗い、chloroformで超音波洗浄(10 min)を行ない、表面を窒素気流で乾燥させ、接触角を測定した。この基板表面の水の接触角は70°であり、基板上にアジド基が導入されたと考えられる。
さらに、50 mL太口ナスフラスコに2-(5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl)propyl N-succinimidyl carbonate 60.5 mg (0.149 mmol)を入れ、窒素気流下においてdry-acetonitrile 10 mL、copper(I) bromide 2.8 mg (19.5 μmol, 0.1eq)を加え、0.5% acetonitrile溶液を調製した。その溶液にアジド化基板を入れ、50 ℃で71時間浸漬した。その後EDTA・Na水溶液で超音波洗浄(10 min)、純水で超音波洗浄(10 min)、methanolで超音波洗浄(10 min)、chloroformで超音波洗浄(10 min)を行ない、表面を窒素気流で乾燥させ、接触角を測定した。この基板表面の水の接触角は64°であり、基板上に光分解性カップリング剤が導入されたことが推測される。
Next, an example in the case of FIG.
First, the silicon wafer substrate was ultrasonically cleaned with H 2 O and methanol, dried with a nitrogen stream, and then pretreated with a UV ozone cleaner. Subsequently, 17.5 mg (85.2 μmol) of (3-azidopropyl) trimethoxysilane was placed in a 50 ml large-mouth eggplant flask, and 4 mL of tetrahydrofuran, 10 μL of 2N HCl, and 16 mL of cyclohexane were added. A pretreated silicon wafer substrate was placed in the solution and immersed for 18.5 hours at room temperature. Then, it was washed with methanol and ultrasonically washed with methanol (10 min). Further, it was washed with chloroform, ultrasonically washed with chloroform (10 min), the surface was dried with a nitrogen stream, and the contact angle was measured. The contact angle of water on the substrate surface is 70 °, and it is considered that an azide group was introduced on the substrate.
Further, add 6-0.5 mg (0.149 mmol) of 2- (5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl) propyl N-succinimidyl carbonate to a 50 mL large-mouth eggplant flask and dry-acetonitrile 10 under nitrogen flow. mL and copper (I) bromide 2.8 mg (19.5 μmol, 0.1 eq) were added to prepare a 0.5% acetonitrile solution. An azide substrate was placed in the solution and immersed at 50 ° C. for 71 hours. Then, ultrasonic cleaning with EDTA / Na aqueous solution (10 min), ultrasonic cleaning with pure water (10 min), ultrasonic cleaning with methanol (10 min), ultrasonic cleaning with chloroform (10 min), nitrogen surface The contact angle was measured by drying with an air stream. The contact angle of water on the substrate surface is 64 °, and it is estimated that a photodegradable coupling agent was introduced onto the substrate.
さらに、上述の工程で得られた光分解性カップリング剤を修飾した基板を用いて、他方の反応基であるスクシンイミジル基に対するアミン化合物の修飾も行った。すなわち、50 mL太口ナスフラスコに窒素雰囲気下において、4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,11-heptadecafluoroundecylamine 31.5 mg (63.6 μmol), dry-acetonitrile 10 mL、triethylamine 4.0 μL (2.9 mg, 28.6 μmol)加えた。その溶液にカップリング剤を修飾した基板を入れ、室温で24時間浸漬した。その後methanolで超音波洗浄(10 min)を行った。さらにchloroformで超音波洗浄(10 min)を行ない、表面を窒素気流で乾燥させ、接触角を測定した(図8)。この基板表面の水の接触角は108°であり、大きく上昇したことから疎水性であるフッ化アルキル鎖が基板上に導入されたことを示す。また、図9に示すXPS測定からもフッ化アルキル鎖が導入されたことを確認した。
その後、得られた修飾基板の光分解を調べるために、超高圧水銀灯で、320 nm以下の波長の光を遮断する硫酸銅フィルターを通して一定時間ごとに光照射し(波長> 320 nm、光量50 mW/ cm2)、水の接触角測定を行った。
光分解は次式のようになされ、図8に示す水の接触角測定より、時間と共に光分解が進行していることが確認された。また、図9に示すXPS測定において、光照射後にフッ化アルキル鎖のピークが消失していることからも光分解を確認した。
The photolysis was performed according to the following equation, and it was confirmed from the measurement of the contact angle of water shown in FIG. Further, in the XPS measurement shown in FIG. 9, the photolysis was also confirmed because the peak of the alkyl fluoride chain disappeared after the light irradiation.
また、上記修飾基板の光分解を調べるために、別の条件での光照射も行った。すなわち、石英セルにアセトニトリル-トリエチルアミン10%溶液中に修飾基板を入れ、超高圧水銀灯で、320 nm以下の波長の光を遮断する硫酸銅フィルターを通して一定時間ごとに光照射し(波長> 320 nm、光量 50 mW/ cm2)、水の接触角測定を行った。
図10に示す水の接触角測定より、空気中で光照射するよりも光分解が促進され、時間と共に光分解が進行していることが確認された。
Moreover, in order to investigate the photodecomposition of the modified substrate, light irradiation was performed under different conditions. That is, a modified substrate is placed in a 10% acetonitrile-triethylamine solution in a quartz cell, and light is irradiated with a super high pressure mercury lamp through a copper sulfate filter that blocks light of a wavelength of 320 nm or less (wavelength> 320 nm, The amount of light was 50 mW / cm 2 ), and the contact angle of water was measured.
From the contact angle measurement of water shown in FIG. 10, it was confirmed that photolysis was promoted more than light irradiation in the air, and that photolysis proceeded with time.
また、光分解性カップリング剤を修飾した基板に対して、上述とは異なるアミン化合物の修飾も行った。すなわち、50 mL太口ナスフラスコに窒素雰囲気下において、PEG350-NH2 22.6 mg (64.8 μmol)、dry-acetonitrile20 mL、triethylamine 10.0 μL (7.26 mg, 71.7 μmol)加え、溶液を調製した。その溶液にカップリング剤を修飾した基板を入れ、室温で22時間攪拌した。その後methanolで超音波洗浄(10 min)を行った。さらにchloroformで超音波洗浄(10 min)を行ない、表面を窒素気流で乾燥させ接触角を測定した。この基板表面の水の接触角は50°であり、減少したことから親水性であるポリエチレングリコール鎖が基板上に導入されたことを示す。
その後、得られた修飾基板の光分解を調べるために、石英セルにアセトニトリル-トリエチルアミン10%溶液中に修飾基板を入れ、超高圧水銀灯で、320 nm以下の波長の光を遮断する硫酸銅フィルターを通して一定時間ごとに光照射し(波長 > 320 nm、光量 50 mW/ cm2)、水の接触角測定を行った。
光分解は次式のようになされ、図11に示す水の接触角測定より、時間と共に光分解が進行していることが確認された。
The photolysis was carried out according to the following equation, and it was confirmed from the measurement of the contact angle of water shown in FIG.
また、このニトロフェネチル型光分解性カップリング剤は下記の式で示されるように、先に片末端に加水分解性シリル基をもつアミンまたはアジドと反応させてから表面に導入することも可能である。
50 mL 二口ナスフラスコに窒素雰囲気下において、2-(5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl)propyl N-succinimidyl carbonate 300 mg (0.738 mmol, 1.0 eq)、dry-tetrahydrofuran 10 mLを入れ、dry-tetrahydrofuran 2 mLに溶解した3-aminopropyltrimethoxysilane 0.147 g (0.818 mmol, 1.1 eq)を加え、室温で2時間攪拌した。濃縮後、中圧カラムクロマトグラフィー(hexane : ethyl acetate : acetone : tetramethoxysilane = 50 : 25 : 25 : 1)、次いでカラムクロマトグラフィー(hexane : ethyl acetate : tetramethoxysilane = 200 : 100 : 3)を行い、黄色粘体185 mg (0.393 mmol)を得た。 2- (5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl) propyl N-succinimidyl carbonate 300 mg (0.738 mmol, 1.0 eq), dry-tetrahydrofuran 10 mL was added, 0.147 g (0.818 mmol, 1.1 eq) of 3-aminopropyltrimethoxysilane dissolved in 2 mL of dry-tetrahydrofuran was added, and the mixture was stirred at room temperature for 2 hours. After concentration, medium pressure column chromatography (hexane: ethyl acetate: acetone: tetramethoxysilane = 50: 25: 25: 1) followed by column chromatography (hexane: ethyl acetate: tetramethoxysilane = 200: 100: 3) 185 mg (0.393 mmol) was obtained.
上記合成で得られた2−(5−メトキシ−2−ニトロ−4−プロパ−2−インイルオキシフェニル)プロピル (3−トリメトキシシリル)プロピルカルバメートの同定結果を以下に示す。
Yield 185 mg (0.393 mmol) 53%
Rf値 0.10 (hexane : ethyl acetate = 2 : 1)
1H-NMR (400 MHz, CDCl3/ TMS)
δ= 0.61 (3H, t, J = 8.1 Hz, -CH2-CH2-CH 2 -Si-)
= 1.33 (2H, d, J = 7.0 Hz, Ar-CH-CH 3 )
= 1.57 - 1.62 (2H, d, J = 7.0 Hz, Ar-CH-CH 3 )
= 2.59 (3H, t, J = 2.4 Hz, CH≡C-)
= 3.12 (2H, q, J = 6.7 Hz, NH-CH 2 )
= 3.56 (9H, s, -Si-(OCH 3 ) 3 )
= 3.89 - 3.95 (4H, m, Ar-OCH 3 , -CH(CH3)-CH2)
= 4.13 - 4.24 (2H, m, -CH(CH3)-CH 2 )
= 4.78 - 4.81 (3H, m, CH≡C-CH 2 -, NH)
= 6.85 (1H, s, Ar-H)
= 7.60 (1H, s, Ar-H)
IR (NaCl)
3340 cm-1 (NH)
3294 cm-1 (C-H alkyne)
2122 cm-1 (C≡C)
1712 cm-1 (C=O)
1520 and 1338 cm-1(-NO2)
The identification results of 2- (5-methoxy-2-nitro-4-prop-2-ynyloxyphenyl) propyl (3-trimethoxysilyl) propyl carbamate obtained by the above synthesis are shown below.
Yield 185 mg (0.393 mmol) 53%
R f value 0.10 (hexane: ethyl acetate = 2: 1)
1 H-NMR (400 MHz, CDCl 3 / TMS)
δ = 0.61 (3H, t, J = 8.1 Hz, -CH 2 -CH 2 -C H 2 -Si-)
= 1.33 (2H, d, J = 7.0 Hz, Ar-CH-C H 3 )
= 1.57-1.62 (2H, d, J = 7.0 Hz, Ar-CH-C H 3 )
= 2.59 (3H, t, J = 2.4 Hz, C H ≡C-)
= 3.12 (2H, q, J = 6.7 Hz, NH-C H 2 )
= 3.56 (9H, s, -Si- (OC H 3 ) 3 )
= 3.89-3.95 (4H, m, Ar-OC H 3 , -C H (CH 3 ) -CH 2 )
= 4.13-4.24 (2H, m, -CH (CH 3 ) -C H 2 )
= 4.78-4.81 (3H, m, CH≡CC H 2- , N H )
= 6.85 (1H, s, Ar-H)
= 7.60 (1H, s, Ar-H)
IR (NaCl)
3340 cm -1 (NH)
3294 cm -1 (CH alkyne)
2122 cm -1 (C≡C)
1712 cm -1 (C = O)
1520 and 1338 cm -1 (-NO 2 )
クマリニルメチル型光分解性シランカップリング剤及びクマリニルメチル型光分解性カップリング剤の実施例を示す。 Examples of coumarinylmethyl type photodegradable silane coupling agent and coumarinylmethyl type photodegradable coupling agent are shown.
このクマリニルメチル型光分解性シランカップリング剤を図12に示す工程により合成した。
先ず、水分離器を取り付けた冷却管を装着した100 mLナスフラスコに4-bromoresorcinol 4.00 g (21.2 mmol, 1.0 eq)、toluene 45 mL、ethyl 4-chloroacetoacetate 5.23 g (31.8 mmol, 1.5 eq)、p-toluenesulfonic acid monohydrate 0.42 g (2.2 mmol, 0.10 eq)を入れ、撹拌しながら14時間加熱還流した。反応混合物を室温まで放冷後、冷水60 mLを加えて30分撹拌した。析出物を吸引ろ過にて回収し、真空乾燥を行い、茶色固体4.32 g (14.9 mmol, 71%)を得た(ステップ7)。
This coumarinylmethyl type photodegradable silane coupling agent was synthesized by the process shown in FIG.
First, in a 100 mL eggplant flask equipped with a condenser equipped with a water separator, 4-bromoresorcinol 4.00 g (21.2 mmol, 1.0 eq), toluene 45 mL, ethyl 4-chloroacetoacetate 5.23 g (31.8 mmol, 1.5 eq), p -toluenesulfonic acid monohydrate 0.42 g (2.2 mmol, 0.10 eq) was added, and the mixture was heated to reflux with stirring for 14 hours. The reaction mixture was allowed to cool to room temperature, 60 mL of cold water was added, and the mixture was stirred for 30 min. The precipitate was collected by suction filtration and vacuum-dried to obtain 4.32 g (14.9 mmol, 71%) of a brown solid (Step 7).
上記合成で得られた6−ブロモ−4−クロロメチル−7−ヒドロキシクマリンの同定結果を以下に示す。
Yield 4.32 g (14.9 mmol, 71%)
Rf = 0.47 (hexane : ethyl acetate = 1 : 1)
1H NMR (CD3OD / TMS) 400 MHz: δ = 4.82 (2H, s, -CH 2 -Cl), 6.42 (1H, s, -CH-C=O), 6.85 (1H, s, -CH-C-OH), 7.95 (1H, s, -CH-C-Br)
IR (KBr): 1703 cm-1(C=O), 3400 cm-1 [Br] (O-H)
The identification result of 6-bromo-4-chloromethyl-7-hydroxycoumarin obtained by the above synthesis is shown below.
Yield 4.32 g (14.9 mmol, 71%)
R f = 0.47 (hexane: ethyl acetate = 1: 1)
1 H NMR (CD 3 OD / TMS) 400 MHz: δ = 4.82 (2H, s, -C H 2 -Cl), 6.42 (1H, s, -C H -C = O), 6.85 (1H, s, -C H -C-OH), 7.95 (1H, s, -C H -C-Br)
IR (KBr): 1703 cm -1 (C = O), 3400 cm -1 [Br] (OH)
次に、10 mLナスフラスコに6-bromo-4-chloromethyl-7-hydroxycoumarin 67.1 mg (0.23 mmol, 1.0 eq)、dry benzene 2 mL、4-pentenoic acid 48 μL (0.47 mmol, 2.0 eq)、DBU 131 μL (0.88 mmol, 3.8 eq)を入れ、N2雰囲気下、1.5時間還流した。ethyl acetateで希釈後、1 N HClでクエンチし、ethyl acetateで抽出、anhydrous MgSO4で乾燥、ろ過、濃縮した。真空乾燥を行い、淡褐色固体71.3 mgを得た。カラムクロマトグラフィー(展開溶媒chloroform : methanol = 20 : 1)、黄色固体65.2 mg (80%)を得た(ステップ8)。 Next, 6-bromo-4-chloromethyl-7-hydroxycoumarin 67.1 mg (0.23 mmol, 1.0 eq), dry benzene 2 mL, 4-pentenoic acid 48 μL (0.47 mmol, 2.0 eq), DBU 131 μL (0.88 mmol, 3.8 eq) was added and refluxed for 1.5 hours under N 2 atmosphere. After diluted with ethyl acetate, quenched with 1 N HCl, extracted with ethyl acetate, dried over anhydrous MgSO 4 , filtered and concentrated. Vacuum drying was performed to obtain 71.3 mg of a light brown solid. Column chromatography (developing solvent: chloroform: methanol = 20: 1), 65.2 mg (80%) of a yellow solid was obtained (step 8).
上記合成で得られた6−ブロモ−7−ヒドロキシクマリン−4−イルメチル 4−ペンテノアートの同定結果を以下に示す。
Yield 65.2 mg (0.18 mmol, 80%)
Rf = 0.33 (hexane : ethyl acetate = 1 : 1), 0.23 (chloroform : methanol = 20 : 1)
1H NMR (CD3OD / TMS) 400 MHz: δ = 2.41 (2H, q, J = 7 Hz, -CH2-CH 2 -CH=CH2), 2.58 (2H, t, J = 7 Hz, -CH 2 -CH2-CH=CH2), 5.00 (1H, d with fine coupling, J = 10 Hz, -CH=CH 2), 5.07 (1H, d with fine coupling, J = 17 Hz, -CH=CH 2), 5.31 (2H, d, J = 1.3 Hz, -CH 2 -O-C(=O)-), 5.86 (1H, ddt, J = 17, 10, 7 Hz, -CH=CH2), 6.26 (1H, t, J = 1.3 Hz, -CH-C=O), 6.85 (1H, s, -CH-C-OH), 7.83 (1H, s, -CH-C-Br)
1H NMR (CDCl3 / TMS) 400 MHz: δ = 2.45 (2H, q, J = 7 Hz, -CH2-CH 2 -CH=CH2), 2.59 (2H, t, J = 7 Hz, -CH 2 -CH2-CH=CH2), 5.06 (1H, d with fine coupling, J = 10 Hz, -CH=CH 2), 5.10 (1H, d with fine coupling, J = 17 Hz, -CH=CH 2), 5.25 (2H, s, -CH 2 -O-C(=O)-), 5.84 (1H, ddt, J = 17, 10, 7 Hz, -CH=CH2), 6.37 (1H, s, -CH-C=O), 7.06 (1H, s, -CH-C-OH), 7.64 (1H, s, -CH-C-Br)
The identification results of 6-bromo-7-hydroxycoumarin-4-ylmethyl 4-pentenoate obtained by the above synthesis are shown below.
Yield 65.2 mg (0.18 mmol, 80%)
R f = 0.33 (hexane: ethyl acetate = 1: 1), 0.23 (chloroform: methanol = 20: 1)
1 H NMR (CD 3 OD / TMS) 400 MHz: δ = 2.41 (2H, q, J = 7 Hz, -CH 2 -C H 2 -CH = CH 2 ), 2.58 (2H, t, J = 7 Hz , -C H 2 -CH 2 -CH = CH 2 ), 5.00 (1H, d with fine coupling, J = 10 Hz, -CH = C H 2 ), 5.07 (1H, d with fine coupling, J = 17 Hz , -CH = C H 2 ), 5.31 (2H, d, J = 1.3 Hz, -C H 2 -OC (= O)-), 5.86 (1H, ddt, J = 17, 10, 7 Hz, -C H = CH 2 ), 6.26 (1H, t, J = 1.3 Hz, -C H -C = O), 6.85 (1H, s, -C H -C-OH), 7.83 (1H, s, -C H -C-Br)
1 H NMR (CDCl 3 / TMS) 400 MHz: δ = 2.45 (2H, q, J = 7 Hz, -CH 2 -C H 2 -CH = CH 2 ), 2.59 (2H, t, J = 7 Hz, -C H 2 -CH 2 -CH = CH 2 ), 5.06 (1H, d with fine coupling, J = 10 Hz, -CH = C H 2 ), 5.10 (1H, d with fine coupling, J = 17 Hz, -CH = C H 2 ), 5.25 (2H, s, -C H 2 -OC (= O)-), 5.84 (1H, ddt, J = 17, 10, 7 Hz, -C H = CH 2 ), 6.37 (1H, s, -C H -C = O), 7.06 (1H, s, -C H -C-OH), 7.64 (1H, s, -C H -C-Br)
次に、20 mL二口ナスフラスコに、6-bromo-7-t-butoxycarbonyloxycoumarin-4-ylmethyl 4-pentenoate 200 mg (0.566 mmol, 1.0 eq)と撹拌子を入れて1時間真空乾燥した後、dry THF 1.0 mL、trimethoxysilane 721 μL (5.66 mmol, 10 eq)、Karstedt’s catalyst 5 drops(パスツールピペット使用、ca. 90 mg)を加え、N2雰囲気下、遮光して室温で16時間撹拌した。エバポレータで濃縮した後、カラムクロマトグラフィーにて精製し(展開溶媒hexane : chloroform : ethyl acetate : tetramethoxysilane = 5 : 5 : 1: 0.1→4 : 4 : 1 : 0.1)、白色固体146 mg (0.308 mmol, 54%)を得た(ステップ9)。 Next, in a 20 mL double-necked eggplant flask, put 6-bromo-7-t-butoxycarbonyloxycoumarin-4-ylmethyl 4-pentenoate 200 mg (0.566 mmol, 1.0 eq) and a stirring bar for 1 hour under vacuum, then dry THF 1.0 mL, trimethoxysilane 721 μL (5.66 mmol, 10 eq) and Karstedt's catalyst 5 drops (using Pasteur pipette, ca. 90 mg) were added, and the mixture was stirred at room temperature for 16 hours under N 2 atmosphere and protected from light. After concentrating with an evaporator, it is purified by column chromatography (developing solvent hexane: chloroform: ethyl acetate: tetramethoxysilane = 5: 5: 1: 0.1 → 4: 4: 1: 0.1), 146 mg (0.308 mmol, 54%) was obtained (step 9).
上記合成で得られた6−ブロモ−7−ヒドロキシクマリン−4−イルメチル 5−トリメトキシシリルペンタノアートの同定結果を以下に示す。
Yield 146 mg (0.308 mmol, 54%)
Rf = 0.37 (hexane : ethyl acetate = 1 : 1)
1H NMR (CDCl3 / TMS) 400 MHz: δ = 0.65-0.71 (2H, m, -CH2-CH2-CH2-CH 2 -Si(OCH3)3), 1.44-1.54 (2H, m, -CH2-CH2-CH 2 -CH2-Si(OCH3)3), 1.74 (2H, quint., J = 7.7 Hz, -CH2-CH 2 -CH2-CH2-Si(OCH3)3), 2.47 (2H, t, J = 7.6 Hz, -CH 2 -CH2-CH2-CH2-Si(OCH3)3), 3.57 (9H, s, -Si-(O-CH 3 ) 3 ), 5.22 (2H, s, -C(=O)-O-CH 2 -), 6.02 (1H, s, -OH), 6.37 (1H, s, -CH-C=O), 7.03 (1H, s, -CH-C-OH), 7.64 (1H, s, -CH-C-Br)
The identification results of 6-bromo-7-hydroxycoumarin-4-ylmethyl 5-trimethoxysilylpentanoate obtained by the above synthesis are shown below.
Yield 146 mg (0.308 mmol, 54%)
R f = 0.37 (hexane: ethyl acetate = 1: 1)
1 H NMR (CDCl 3 / TMS) 400 MHz: δ = 0.65-0.71 (2H, m, -CH 2 -CH 2 -CH 2 -C H 2 -Si (OCH 3 ) 3 ), 1.44-1.54 (2H, m, -CH 2 -CH 2 -C H 2 -CH 2 -Si (OCH 3 ) 3 ), 1.74 (2H, quint., J = 7.7 Hz, -CH 2 -C H 2 -CH 2 -CH 2- Si (OCH 3 ) 3 ), 2.47 (2H, t, J = 7.6 Hz, -C H 2 -CH 2 -CH 2 -CH 2 -Si (OCH 3 ) 3 ), 3.57 (9H, s, -Si- (OC H 3 ) 3 ), 5.22 (2H, s, -C (= O) -OC H 2- ), 6.02 (1H, s, -O H), 6.37 (1H, s, -C H -C = O), 7.03 (1H, s, -C H -C-OH), 7.64 (1H, s, -C H -C-Br)
以上の工程で得られたクマリニルメチル型光分解性シランカップリング剤を用いて表面修飾を行う場合の例を図13に示す。
先ず、50 mL太口ナスフラスコにconc. H2SO4 (14 mL)と30% H2O2 (6 mL)を入れて混合し、ピラニア溶液(conc. H2SO4: 30% H2O2 = 7 : 3)20 mLを調製した。シリコンウェハー基板を入れ、100 ℃で1 h静置した。室温まで放冷後、溶液を除去して、純水約20 mLで3回洗浄し、純水約30 mL中で10 min超音波洗浄した。窒素気流で乾燥し、表面修飾に用いた。
続いて、50 mL太口ナスフラスコに6-bromo-7-hydroxycoumarin-4-ylmethyl 5-trimethoxysilylpentanoate (9.5 mg, 0.02 mmol)の1 mM dry toluene溶液20 mLを調製し、シリコンウェハー基板を入れ、N2雰囲気下、遮光して室温で3 h静置した。基板を取り出し、methanolでリンスし、methanol約30 mL中で10 min超音波洗浄し、次いでchloroformでリンスし、chloroform約30 mL中で10 min超音波洗浄し、窒素気流で乾燥した。この基板表面の水の接触角は61°であり、基板上にクマリニルメチル型光分解性シランカップリング剤が導入されたことが推測される。
FIG. 13 shows an example in which surface modification is performed using the coumarinylmethyl type photodegradable silane coupling agent obtained in the above steps.
First, conc. H 2 SO 4 (14 mL) and 30% H 2 O 2 (6 mL) are placed in a 50 mL large-mouth eggplant flask and mixed to form a piranha solution (conc. H 2 SO 4 : 30% H 2 O 2 = 7: 3) 20 mL was prepared. A silicon wafer substrate was placed and allowed to stand at 100 ° C. for 1 h. After allowing to cool to room temperature, the solution was removed, washed with about 20 mL of pure water three times, and ultrasonically washed for 10 min in about 30 mL of pure water. It dried with nitrogen stream and used for surface modification.
Subsequently, 20 mL of a 1 mM dry toluene solution of 6-bromo-7-hydroxycoumarin-4-ylmethyl 5-trimethoxysilylpentanoate (9.5 mg, 0.02 mmol) was prepared in a 50 mL large-mouth eggplant flask. The mixture was allowed to stand for 3 hours at room temperature in the dark and protected from light. The substrate was taken out, rinsed with methanol, ultrasonically cleaned in about 30 mL of methanol, then rinsed with chloroform, ultrasonically cleaned in about 30 mL of chloroform, and dried in a nitrogen stream. The contact angle of water on the substrate surface is 61 °, and it is estimated that a coumarinylmethyl type photodegradable silane coupling agent was introduced onto the substrate.
その後、6−ブロモ−7−ヒドロキシクマリン−4−イルメチル 5−トリメトキシシリルペンタノアートを表面に導入した修飾基板の光分解を調べるために、超高圧水銀灯で、320 nm以下の波長の光を遮断する硫酸銅フィルターを通して1% DMSO-H2O溶液中で一定時間ごとに光照射し(波長 > 320 nm、光量15 mW/ cm2)、水の接触角測定を行った。
光分解は次式のようになされ、図14に示す水の接触角測定より、時間と共に光分解が進行していることが確認された。また、図15に示すXPS測定において、光照射後にブロモ基由来のピークが消失していることからも光分解を確認した。
Then, in order to investigate the photolysis of the modified substrate into which 6-bromo-7-hydroxycoumarin-4-ylmethyl 5-trimethoxysilylpentanoate was introduced on the surface, light with a wavelength of 320 nm or less was irradiated with an ultrahigh pressure mercury lamp. The contact angle of water was measured by irradiating light in a 1% DMSO-H 2 O solution at regular intervals (wavelength> 320 nm, light amount 15 mW / cm 2 ) through a copper sulfate filter to be blocked.
The photolysis was performed as follows, and it was confirmed from the measurement of the contact angle of water shown in FIG. Further, in the XPS measurement shown in FIG. 15, photolysis was also confirmed from the disappearance of the peak derived from the bromo group after the light irradiation.
また、クマリニルメチル型光分解性カップリング剤を図16に示す工程により合成した。
先ず、200 mLナスフラスコに6-bromo-4-chloromethyl-7-hydroxycoumarin 2.03 g (7.01 mmol, 1.0 eq)、DMF 60 mL、1 N HCl 30 mLを入れ、100 °Cで19時間(終夜)撹拌した。減圧蒸留によりDMFを除去し、残渣にsat. NaCl aq. 50 mLを加え、ethyl acetate (100 mL x 2, 50 mL x 3)で抽出し、有機層をanhydrous MgSO4で乾燥、ろ過、濃縮した。真空乾燥(湯浴45 °C)を行い、茶色固体1.68 g (6.20 mmol, 88%)を得た(ステップ10)。
A coumarinylmethyl-type photodegradable coupling agent was synthesized by the process shown in FIG.
First, add 2.03 g (7.01 mmol, 1.0 eq) of 6-bromo-4-chloromethyl-7-hydroxycoumarin, 60 mL of DMF, and 30 mL of 1 N HCl to a 200 mL eggplant flask and stir at 100 ° C for 19 hours (all night) did. Remove DMF by vacuum distillation, add sat. NaCl aq. 50 mL to the residue, extract with ethyl acetate (100 mL x 2, 50 mL x 3), dry the organic layer with anhydrous MgSO 4 , filter and concentrate . Vacuum drying (water bath 45 ° C) was performed to obtain 1.68 g (6.20 mmol, 88%) of a brown solid (Step 10).
上記合成で得られた6−ブロモ−7−ヒドロキシ−4−ヒドロキシメチルクマリンの同定結果を以下に示す。
Yield 1.68 g (6.20 mmol, 88%)
Rf = 0.18 (hexane : ethyl acetate = 1 : 1)
1H NMR (CD3OD / TMS) 400 MHz: δ = 4.77 (2H, d, J= 1.4 Hz, -CH 2 -OH), 6.38 (1H, t, J = 1.4 Hz, -CH-C=O), 6.84 (1H, s, -CH-C-OH), 7.82 (1H, s, -CH-C-Br)
The identification results of 6-bromo-7-hydroxy-4-hydroxymethylcoumarin obtained by the above synthesis are shown below.
Yield 1.68 g (6.20 mmol, 88%)
R f = 0.18 (hexane: ethyl acetate = 1: 1)
1 H NMR (CD 3 OD / TMS) 400 MHz: δ = 4.77 (2H, d, J = 1.4 Hz, -C H 2 -OH), 6.38 (1H, t, J = 1.4 Hz, -C H -C = O), 6.84 (1H, s, -C H -C-OH), 7.82 (1H, s, -C H -C-Br)
次に、20 mL二口ナスフラスコに、6-bromo-7-hydroxy-4-hydroxymethylcoumarin 200 mg (0.74 mmol, 1.0 eq)、DMAP 55.8 mg (0.46 mmol, 0.6 eq)、EDC・HCl 177 mg (0.92 mmol, 1.3 eq)、dry THF 4 mL、5-hexynoic acid 98 μL (0.89 mmol, 1.2 eq)を入れ、N2雰囲気下、遮光して室温で5時間撹拌した。1 N HCl (5 mL)を加えてクエンチし、ethyl acetate (5 mL x 3)で抽出、有機層をsat. NaCl aq. (5 mL)で洗浄後、anhydrous MgSO4で乾燥、ろ過、濃縮した。真空乾燥を行い、茶色粘体118 mgを得た。カラムクロマトグラフィーにて精製し(展開溶媒chloroform : ethyl acetate = 15 : 1)、淡茶色固体86 mg (32%)を得た(ステップ11)。 Next, in a 20 mL two-necked eggplant flask, 6-bromo-7-hydroxy-4-hydroxymethylcoumarin 200 mg (0.74 mmol, 1.0 eq), DMAP 55.8 mg (0.46 mmol, 0.6 eq), EDC / HCl 177 mg (0.92 mmol, 1.3 eq), dry THF 4 mL, 5-hexynoic acid 98 μL (0.89 mmol, 1.2 eq) was added, and the mixture was stirred at room temperature for 5 hours in a N 2 atmosphere, protected from light. 1 N was quenched by addition of HCl (5 mL), extracted with ethyl acetate (5 mL x 3) , the organic layer was washed sat. NaCl aq. At (5 mL), dried over Anhydrous MgSO 4, filtered, and concentrated . Vacuum drying was performed to obtain 118 mg of a brown viscous body. Purification by column chromatography (developing solvent chloroform: ethyl acetate = 15: 1) gave 86 mg (32%) of a light brown solid (step 11).
上記合成で得られた6−ブロモ−7−ヒドロキシクマリン−4−イルメチル 5−ヘキシノアートの同定結果を以下に示す。
Yield 86 mg (0.23 mmol, 32%)
Rf = 0.28 (chloroform : ethyl acetate = 15 : 1)
1H NMR (CDCl3 / TMS) 400 MHz: δ = 1.92 (2H, quint., J = 7.2 Hz, -CH2-CH 2 -CH2-C≡CH), 2.01 (1H, t, J = 2.7 Hz, -CH2-CH2-CH2-C≡CH), 2.32 (2H, td, J = 6.9, 2.7 Hz, -CH2-CH2-CH 2 -C≡CH), 2.63 (2H, t, J = 7.5 Hz, -CH 2 -CH2-CH2-C≡CH), 5.23 (2H, d, J = 1.4 Hz, -CH 2 -O-C(=O)-), 6.00 (iH, br s, -OH), 6.38 (1H, t, J= 1.4 Hz, -CH-C=O), 7.03 (1H, s, -CH-C-OH), 7.64 (1H, s, -CH-C-Br)
The identification result of 6-bromo-7-hydroxycoumarin-4-ylmethyl 5-hexinoate obtained by the above synthesis is shown below.
Yield 86 mg (0.23 mmol, 32%)
R f = 0.28 (chloroform: ethyl acetate = 15: 1)
1 H NMR (CDCl 3 / TMS) 400 MHz: δ = 1.92 (2H, quint., J = 7.2 Hz, -CH 2 -C H 2 -CH 2 -C≡CH), 2.01 (1H, t, J = 2.7 Hz, -CH 2 -CH 2 -CH 2 -C≡C H ), 2.32 (2H, td, J = 6.9, 2.7 Hz, -CH 2 -CH 2 -C H 2 -C≡CH), 2.63 ( 2H, t, J = 7.5 Hz , - CH 2 -CH 2 -CH 2 -C≡CH), 5.23 (2H, d, J = 1.4 Hz, -C H 2 -OC (= O) -), 6.00 ( iH, br s, -O H ), 6.38 (1H, t, J = 1.4 Hz, -C H -C = O), 7.03 (1H, s, -C H -C-OH), 7.64 (1H, s , -C H -C-Br)
以上の工程で得られたクマリニルメチル型光分解性カップリング剤である6−ブロモ−7−ヒドロキシクマリン−4−イルメチル 5−ヘキシノアートは、ステップ7で得られた6−ブロモ−4−クロロメチル−7−ヒドロキシクマリンから1段階で合成することもできる。
すなわち、100 mL二口ナスフラスコに、6-bromo-4-chloromethyl-7-hydroxycoumarin 0.500 g (1.73 mmol, 1.0 eq)、potassium carbonate 0.371 g (2.68 mmol, 1.6 eq)、5-hexynoic acid 0.311 g (2.77 mmol, 1.6 eq) 、dry DMF 12.5 mLを入れ、N2雰囲気下80 ℃で2時間加熱した。反応溶液にethyl acetate (25 mL x 3)、1 N HCl 25 mLを加えて抽出し、有機層をsat. NaCl aq. (50 mL x 5)で洗浄、anhydrous MgSO4で乾燥、ろ過、濃縮した。カラムクロマトグラフィーにて精製し(展開溶媒chloroform : methanol = 20 : 1)、淡黄色固体444 mg (1.22 mmol, 70%)を得た(ステップ12)。
6-Bromo-7-hydroxycoumarin-4-ylmethyl 5-hexinoate, which is a coumarinylmethyl-type photodegradable coupling agent obtained in the above process, is obtained from 6-bromo-4-chloromethyl obtained in Step 7. It can also be synthesized in one step from -7-hydroxycoumarin.
That is, in a 100 mL double neck eggplant flask, 6-bromo-4-chloromethyl-7-hydroxycoumarin 0.500 g (1.73 mmol, 1.0 eq), potassium carbonate 0.371 g (2.68 mmol, 1.6 eq), 5-hexynoic acid 0.311 g ( 2.77 mmol, 1.6 eq) and 12.5 mL of dry DMF were added and heated at 80 ° C. for 2 hours under N 2 atmosphere. The reaction solution was extracted with ethyl acetate (25 mL x 3) and 1 N HCl (25 mL), and the organic layer was washed with sat. NaCl aq. (50 mL x 5), dried over anhydrous MgSO 4 , filtered, and concentrated. . Purification by column chromatography (developing solvent chloroform: methanol = 20: 1) gave 444 mg (1.22 mmol, 70%) of a pale yellow solid (step 12).
以上の工程で得られたクマリニルメチル型光分解性カップリング剤を用いて表面修飾を行う場合の例を図17に示す。アルキン反応性基(アジド基)を含むシランカップリング剤を用いて基板表面上に単分子膜を形成し、このアジド基に上述したクマリニルメチル型光分解性カップリング剤の反応基(アルキニル基)を結合させるものである。 FIG. 17 shows an example of surface modification using the coumarinylmethyl type photodegradable coupling agent obtained in the above steps. A monomolecular film is formed on the substrate surface using a silane coupling agent containing an alkyne reactive group (azide group), and the above-described reactive group (alkynyl group) of the coumarinylmethyl type photodegradable coupling agent is formed on this azide group. ).
先ず、シリコンウェハー基板をピラニア溶液(conc. H2SO4: 30% H2O2 = 7 : 3)中、100 ℃で1 h前処理した。次いで、50 mL太口ナスフラスコに(3-azidoproply)trimethoxysilane 18.3 mg (89.1 μmol)、tetrahydrofuran 4 mL、2N HCl 10 μL、cyclohexane 16 mLを入れて溶液を調製した。その溶液に前処理済みシリコンウェハー基板を室温で22 h浸漬した。methanolでリンスし、methanolで10 min超音波洗浄し、次いでchloroformでリンスし、chloroformで10 min超音波洗浄した。表面を窒素気流で乾燥した。この基板表面の水の接触角は75°であり、基板上にアジド基が導入されたと考えられる。
続いて、50 mL太口ナスフラスコに6-bromo-7-hydroxycoumarin-4-ylmethyl 5-hexynoate 55.3 mg (0.151 mmol)を入れ、窒素気流下においてdry-acetonitrile 15 mL、copper(I) bromide 33.4 mg (0.233 mmol, 1.2 eq)を加え、acetonitrile溶液を調製した。その溶液にアジド化基板を入れ、撹拌下50 ℃で24時間浸漬した。その後methanolで10分間超音波洗浄、EDTA・Na aq.で10分間超音波洗浄、純水で15分間超音波洗浄、methanolで10分間超音波洗浄、chloroformで10分間超音波洗浄し、窒素気流で乾燥した。この基板表面の水の接触角は63°であり、基板上に光分解性カップリング剤が導入されたことが推測される。
First, a silicon wafer substrate was pretreated at 100 ° C. for 1 h in a piranha solution (conc. H 2 SO 4 : 30% H 2 O 2 = 7: 3). Subsequently, (3-azidoproply) trimethoxysilane 18.3 mg (89.1 μmol), tetrahydrofuran 4 mL, 2N HCl 10 μL, and cyclohexane 16 mL were prepared in a 50 mL large-mouth eggplant flask. A pretreated silicon wafer substrate was immersed in the solution for 22 hours at room temperature. Rinse with methanol, ultrasonic wash with methanol for 10 min, then rinse with chloroform and ultrasonic wash with chloroform for 10 min. The surface was dried with a nitrogen stream. The contact angle of water on the substrate surface was 75 °, and it is considered that an azide group was introduced onto the substrate.
Subsequently, 6-bromo-7-hydroxycoumarin-4-ylmethyl 5-hexynoate 55.3 mg (0.151 mmol) was placed in a 50 mL large-mouth eggplant flask, and dry-acetonitrile 15 mL, copper (I) bromide 33.4 mg under a nitrogen stream (0.233 mmol, 1.2 eq) was added to prepare an acetonitrile solution. The azide substrate was put into the solution and immersed for 24 hours at 50 ° C. with stirring. Then ultrasonic wash with methanol for 10 minutes, ultrasonic wash with EDTA / Na aq. For 10 minutes, ultrasonic wash with pure water for 15 minutes, ultrasonic wash with methanol for 10 minutes, ultrasonic wash with chloroform for 10 minutes, and nitrogen flow Dried. The contact angle of water on the surface of the substrate is 63 °, and it is assumed that a photodegradable coupling agent was introduced onto the substrate.
その後、6−ブロモ−7−ヒドロキシクマリン−4−イルメチル 5−ヘキシノアートを表面に導入した修飾基板の光分解を調べるために、超高圧水銀灯で、320 nm以下の波長の光を遮断する硫酸銅フィルターを通して1% DMSO-H2O溶液中で一定時間ごとに光照射し(波長 > 320 nm、光量15 mW/ cm2)、水の接触角測定を行った。
光分解は次式のようになされ、図18に示す水の接触角測定より、時間と共に光分解が進行していることが確認された。また、図19に示すXPS測定において、光照射後にブロモ基由来のピークがほぼ消失していることからも光分解を確認した。
Then, in order to investigate the photodecomposition of the modified substrate into which 6-bromo-7-hydroxycoumarin-4-ylmethyl 5-hexinoate is introduced on the surface, a copper sulfate filter that blocks light with a wavelength of 320 nm or less with an ultrahigh pressure mercury lamp. Through a 1% DMSO-H 2 O solution, light was irradiated at regular intervals (wavelength> 320 nm, light intensity 15 mW / cm 2 ), and the contact angle of water was measured.
Photolysis was performed as follows, and it was confirmed from the measurement of the contact angle of water shown in FIG. Further, in the XPS measurement shown in FIG. 19, the photodegradation was also confirmed from the fact that the peak derived from the bromo group almost disappeared after the light irradiation.
尚、以上の光分解性カップリング剤は、特開2007−186472号公報に示される光分解性カップリング剤のスペーサR(光分解性基)として用いるようにしても、また、特開2010−260831号公報に示される光分解性ヘテロ二価性架橋剤のスペーサR(光分解性基)等として用いるようにしてもよい。
即ち、下記一般式(4)で表されるヘテロ二価性の架橋剤(光分解性カップリング剤)として、無機材料と反応する加水分解性を有するシリル基(加水分解性シリル基)、生体分子などのアミノ基と反応するアミン反応性基、及び生体分子などのチオール基と反応するチオール反応性基の中から選択された2つの異なる反応性基X,Yを有し、この2つの異なる反応性基の間に上述したリンカー(ニトロフェネチル型光分解性基またはクマリニルメチル型光分解性基):Rを用いるようにしてもよい。
X−R−Y (4)
The above photodegradable coupling agent may be used as a spacer R (photodegradable group) of a photodegradable coupling agent disclosed in JP-A-2007-186472, or JP-A-2010-. You may make it use as spacer R (photodegradable group) etc. of the photodegradable heterobivalent crosslinking agent shown by 260831 gazette.
That is, as a heterobivalent crosslinking agent (photodegradable coupling agent) represented by the following general formula (4), a hydrolyzable silyl group (hydrolyzable silyl group) that reacts with an inorganic material, It has two different reactive groups X, Y selected from among amine reactive groups that react with amino groups such as molecules and thiol reactive groups that react with thiol groups such as biomolecules. The aforementioned linker (nitrophenethyl-type photodegradable group or coumarinylmethyl-type photodegradable group): R may be used between the reactive groups.
X-R-Y (4)
加水分解性シリル基は、加水分解して無機材料(ガラス、シリコンなど)と結合するものであり、アルコール型、クロロ型、酢酸型、オキシム型、アミド型、アセトン型などが利用可能であり、中でもクロロシリル基またはアルコキシシリル基が好ましい。 Hydrolyzable silyl groups are those that hydrolyze and bond to inorganic materials (glass, silicon, etc.), and are available in alcohol, chloro, acetic, oxime, amide, and acetone types. Of these, a chlorosilyl group or an alkoxysilyl group is preferred.
アミン反応性基は、生体分子等にあるアミン官能基と共有結合するものであり、N-ヒドロキシスクシンイミド( NHS )エステル基、イミドエステル基、イソシアネート基、ニトロフェニルハライド基などが利用可能であり、好適な例としては、下記の構造式(5)で示されるN-ヒドロキシスクシンイミドエステル基を含有するものである。 The amine reactive group is covalently bonded to an amine functional group in a biomolecule or the like, and N-hydroxysuccinimide (NHS) ester group, imide ester group, isocyanate group, nitrophenyl halide group, etc. can be used. Preferable examples include those containing an N-hydroxysuccinimide ester group represented by the following structural formula (5).
チオール反応性基は、生体分子等にあるチオール官能基と共有結合するものであり、マレイミド、ピリジルジスルフィド、チオフタルイミド、活性ハロゲン、ハロアセトアミド、ジスルフィド、チオサルフェートなどが利用可能であり、好適な例としては、下記の構造式(6)又は(7)で示される マレイミドを包含するものである。尚、構造式(6)はチオール反応性基であるが、構造式(7)は加熱によって(6)を生成するチオール反応性基の前駆体である。 The thiol-reactive group is a covalent bond with a thiol functional group in a biomolecule or the like, and maleimide, pyridyl disulfide, thiophthalimide, active halogen, haloacetamide, disulfide, thiosulfate, etc. can be used. Includes a maleimide represented by the following structural formula (6) or (7). Structural formula (6) is a thiol-reactive group, but structural formula (7) is a precursor of a thiol-reactive group that generates (6) by heating.
Claims (2)
一般式(1)
General formula (1)
一般式(2)
General formula (2)
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