JP2014111562A - Sugar derivatives and antimicrobial agents using the same - Google Patents
Sugar derivatives and antimicrobial agents using the same Download PDFInfo
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- JP2014111562A JP2014111562A JP2013165760A JP2013165760A JP2014111562A JP 2014111562 A JP2014111562 A JP 2014111562A JP 2013165760 A JP2013165760 A JP 2013165760A JP 2013165760 A JP2013165760 A JP 2013165760A JP 2014111562 A JP2014111562 A JP 2014111562A
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- Prior art keywords
- group
- salt
- monosaccharide
- azide
- sugar derivative
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 235000000346 sugar Nutrition 0.000 title claims description 26
- 239000004599 antimicrobial Substances 0.000 title abstract 2
- 150000003839 salts Chemical class 0.000 claims abstract description 22
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 18
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 10
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 claims abstract description 10
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 7
- 125000002947 alkylene group Chemical group 0.000 claims abstract description 6
- 125000006176 2-ethylbutyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(C([H])([H])*)C([H])([H])C([H])([H])[H] 0.000 claims abstract description 5
- 125000004850 cyclobutylmethyl group Chemical group C1(CCC1)C* 0.000 claims abstract description 5
- 125000004210 cyclohexylmethyl group Chemical group [H]C([H])(*)C1([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C1([H])[H] 0.000 claims abstract description 5
- 125000004851 cyclopentylmethyl group Chemical group C1(CCCC1)C* 0.000 claims abstract description 5
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 claims abstract description 5
- 150000002772 monosaccharides Chemical class 0.000 claims description 25
- 239000003242 anti bacterial agent Substances 0.000 claims description 24
- 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 claims description 21
- 239000008103 glucose Substances 0.000 claims description 21
- 229920001542 oligosaccharide Polymers 0.000 claims description 16
- 150000002482 oligosaccharides Chemical class 0.000 claims description 16
- 150000001540 azides Chemical class 0.000 claims description 15
- 150000001345 alkine derivatives Chemical group 0.000 claims description 14
- 125000004122 cyclic group Chemical group 0.000 claims description 14
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 7
- -1 saccharide azide Chemical class 0.000 claims description 7
- 230000002194 synthesizing effect Effects 0.000 claims description 7
- IVRMZWNICZWHMI-UHFFFAOYSA-N azide group Chemical group [N-]=[N+]=[N-] IVRMZWNICZWHMI-UHFFFAOYSA-N 0.000 claims description 6
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 claims description 6
- RUPAXCPQAAOIPB-UHFFFAOYSA-N tert-butyl formate Chemical group CC(C)(C)OC=O RUPAXCPQAAOIPB-UHFFFAOYSA-N 0.000 claims description 6
- 239000004480 active ingredient Substances 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 5
- 125000004432 carbon atom Chemical group C* 0.000 claims description 4
- 150000002402 hexoses Chemical class 0.000 claims description 4
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 4
- 125000001424 substituent group Chemical group 0.000 claims description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 3
- 239000001257 hydrogen Substances 0.000 claims description 3
- 229910052739 hydrogen Inorganic materials 0.000 claims description 3
- 125000001425 triazolyl group Chemical group 0.000 claims 1
- 230000015572 biosynthetic process Effects 0.000 abstract description 11
- 238000003786 synthesis reaction Methods 0.000 abstract description 10
- 230000000845 anti-microbial effect Effects 0.000 abstract 1
- 230000008878 coupling Effects 0.000 abstract 1
- 238000010168 coupling process Methods 0.000 abstract 1
- 238000005859 coupling reaction Methods 0.000 abstract 1
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 54
- 230000000844 anti-bacterial effect Effects 0.000 description 40
- 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 39
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 33
- 150000001875 compounds Chemical class 0.000 description 32
- 229920000858 Cyclodextrin Polymers 0.000 description 30
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 30
- 239000000126 substance Substances 0.000 description 26
- GDSRMADSINPKSL-HSEONFRVSA-N gamma-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO GDSRMADSINPKSL-HSEONFRVSA-N 0.000 description 22
- 235000001727 glucose Nutrition 0.000 description 21
- 238000005481 NMR spectroscopy Methods 0.000 description 20
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 18
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 17
- 241000894006 Bacteria Species 0.000 description 16
- 239000007787 solid Substances 0.000 description 16
- 238000006243 chemical reaction Methods 0.000 description 14
- 239000000243 solution Substances 0.000 description 13
- 150000003852 triazoles Chemical group 0.000 description 13
- 229920001282 polysaccharide Polymers 0.000 description 12
- 239000005017 polysaccharide Substances 0.000 description 12
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 11
- 229940080345 gamma-cyclodextrin Drugs 0.000 description 11
- 150000004676 glycans Chemical class 0.000 description 11
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 10
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 description 10
- 239000012528 membrane Substances 0.000 description 10
- 239000002904 solvent Substances 0.000 description 10
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 9
- 238000012650 click reaction Methods 0.000 description 9
- 238000010511 deprotection reaction Methods 0.000 description 9
- 238000004821 distillation Methods 0.000 description 9
- 239000000203 mixture Substances 0.000 description 9
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 8
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 8
- 239000003729 cation exchange resin Substances 0.000 description 8
- 230000012010 growth Effects 0.000 description 8
- 241000588724 Escherichia coli Species 0.000 description 7
- 125000003277 amino group Chemical group 0.000 description 7
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 6
- 238000005873 Huisgen reaction Methods 0.000 description 6
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 6
- 241000191967 Staphylococcus aureus Species 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 229910000365 copper sulfate Inorganic materials 0.000 description 6
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 6
- 238000013461 design Methods 0.000 description 6
- 238000001914 filtration Methods 0.000 description 6
- 235000010378 sodium ascorbate Nutrition 0.000 description 6
- PPASLZSBLFJQEF-RKJRWTFHSA-M sodium ascorbate Substances [Na+].OC[C@@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RKJRWTFHSA-M 0.000 description 6
- 229960005055 sodium ascorbate Drugs 0.000 description 6
- PPASLZSBLFJQEF-RXSVEWSESA-M sodium-L-ascorbate Chemical compound [Na+].OC[C@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RXSVEWSESA-M 0.000 description 6
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 5
- 229910021529 ammonia Inorganic materials 0.000 description 5
- 239000011734 sodium Substances 0.000 description 5
- 244000063299 Bacillus subtilis Species 0.000 description 4
- 235000014469 Bacillus subtilis Nutrition 0.000 description 4
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 4
- 241000192125 Firmicutes Species 0.000 description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 4
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 4
- 239000007795 chemical reaction product Substances 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 150000002304 glucoses Chemical class 0.000 description 4
- 230000002209 hydrophobic effect Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 108010093965 Polymyxin B Proteins 0.000 description 3
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 3
- 239000012300 argon atmosphere Substances 0.000 description 3
- 125000004744 butyloxycarbonyl group Chemical group 0.000 description 3
- 125000002091 cationic group Chemical group 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 229920000024 polymyxin B Polymers 0.000 description 3
- 229960005266 polymyxin b Drugs 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- LXAHHHIGZXPRKQ-UHFFFAOYSA-N 5-fluoro-2-methylpyridine Chemical compound CC1=CC=C(F)C=N1 LXAHHHIGZXPRKQ-UHFFFAOYSA-N 0.000 description 2
- RKWCGBZOIJEXQO-UHFFFAOYSA-N C1(CCC1)CNC#CC Chemical compound C1(CCC1)CNC#CC RKWCGBZOIJEXQO-UHFFFAOYSA-N 0.000 description 2
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 2
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 description 2
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 2
- 229930091371 Fructose Natural products 0.000 description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- 239000005715 Fructose Substances 0.000 description 2
- 229920001503 Glucan Polymers 0.000 description 2
- 229920000057 Mannan Polymers 0.000 description 2
- 239000012359 Methanesulfonyl chloride Substances 0.000 description 2
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 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 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 229910052802 copper Inorganic materials 0.000 description 2
- 239000010949 copper Substances 0.000 description 2
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 description 2
- 239000012039 electrophile Substances 0.000 description 2
- 229930182830 galactose Natural products 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 125000001165 hydrophobic group Chemical group 0.000 description 2
- QARBMVPHQWIHKH-UHFFFAOYSA-N methanesulfonyl chloride Chemical compound CS(Cl)(=O)=O QARBMVPHQWIHKH-UHFFFAOYSA-N 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 239000012038 nucleophile Substances 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 238000001308 synthesis method Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- ODPLTKNMIDPDKY-UHFFFAOYSA-N 2-methyl-N-prop-1-ynylpropan-1-amine Chemical compound CC#CNCC(C)C ODPLTKNMIDPDKY-UHFFFAOYSA-N 0.000 description 1
- OVYJFLKSSYXZOT-UHFFFAOYSA-N 2-methyl-n-(2h-triazol-4-ylmethyl)propan-1-amine Chemical group CC(C)CNCC=1C=NNN=1 OVYJFLKSSYXZOT-UHFFFAOYSA-N 0.000 description 1
- JSIAIROWMJGMQZ-UHFFFAOYSA-N 2h-triazol-4-amine Chemical group NC1=CNN=N1 JSIAIROWMJGMQZ-UHFFFAOYSA-N 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 229920000856 Amylose Polymers 0.000 description 1
- VLHBJGRTAKWQTO-UHFFFAOYSA-N C(C)C(CNC#CC)CC Chemical compound C(C)C(CNC#CC)CC VLHBJGRTAKWQTO-UHFFFAOYSA-N 0.000 description 1
- VTSWFYPSKKQQOG-UHFFFAOYSA-N C(C)C(CNCC=1N=NNC1)CC Chemical group C(C)C(CNCC=1N=NNC1)CC VTSWFYPSKKQQOG-UHFFFAOYSA-N 0.000 description 1
- FIHOHTRQBHEMTG-UHFFFAOYSA-N C1(CCCC1)CNC#CC Chemical compound C1(CCCC1)CNC#CC FIHOHTRQBHEMTG-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 108010069514 Cyclic Peptides Proteins 0.000 description 1
- 102000001189 Cyclic Peptides Human genes 0.000 description 1
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- RJQXTJLFIWVMTO-TYNCELHUSA-N Methicillin Chemical compound COC1=CC=CC(OC)=C1C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 RJQXTJLFIWVMTO-TYNCELHUSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical group OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 125000004202 aminomethyl group Chemical group [H]N([H])C([H])([H])* 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 229940124350 antibacterial drug Drugs 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 125000001995 cyclobutyl group Chemical group [H]C1([H])C([H])([H])C([H])(*)C1([H])[H] 0.000 description 1
- 229940097362 cyclodextrins Drugs 0.000 description 1
- 125000000113 cyclohexyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 150000002303 glucose derivatives Chemical class 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229920006008 lipopolysaccharide Polymers 0.000 description 1
- LUEWUZLMQUOBSB-GFVSVBBRSA-N mannan Chemical class 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]2[C@H](O[C@@H](O[C@H]3[C@H](O[C@@H](O)[C@@H](O)[C@H]3O)CO)[C@@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-GFVSVBBRSA-N 0.000 description 1
- 229960003085 meticillin Drugs 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- SRWFRXQZAMQQOS-UHFFFAOYSA-N n-(cyclohexylmethyl)prop-2-yn-1-amine Chemical compound C#CCNCC1CCCCC1 SRWFRXQZAMQQOS-UHFFFAOYSA-N 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 125000000467 secondary amino group Chemical group [H]N([*:1])[*:2] 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000000707 stereoselective effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Saccharide Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
Description
本発明は、糖誘導体及びそれを用いた抗菌剤に関する。 The present invention relates to a sugar derivative and an antibacterial agent using the sugar derivative.
ペニシリンの発見以来、様々な抗生物質や合成抗菌薬が開発され、感染症治療のために使用されてきた。しかし、メシチリン耐性黄色ブドウ球菌（ＭＲＳＡ）に代表される既存の抗菌薬に対して耐性を持つ多剤耐性菌が深刻なまでに増加し、新たな抗菌薬の発展が渇望されている。 Since the discovery of penicillin, various antibiotics and synthetic antibacterial drugs have been developed and used to treat infections. However, multidrug-resistant bacteria having resistance to existing antibacterial agents typified by methicillin-resistant Staphylococcus aureus (MRSA) have been seriously increased, and the development of new antibacterial agents is eagerly desired.
一般に薬剤耐性が起こりにくい抗菌物質として、ポリミキシンBなどの膜作動型機構により抗菌性を示す物質が存在する。しかし、これらは複雑な構造を有する天然物であり、単離したり、人工合成したりすることは困難である。このため、人工的な抗菌性物質を合成することも行われており、特異な化学構造を有するシクロデキストリンを化学修飾して抗菌剤として用いることが行われている（例えば特許文献１、２）。 In general, as antibacterial substances that hardly cause drug resistance, there are substances that exhibit antibacterial properties by a membrane-actuated mechanism such as polymyxin B. However, these are natural products having a complicated structure and are difficult to isolate or artificially synthesize. For this reason, an artificial antibacterial substance is also synthesized, and cyclodextrin having a specific chemical structure is chemically modified and used as an antibacterial agent (for example, Patent Documents 1 and 2). .
抗菌作用のメカニズムが詳細には解明されていないことが多く、人工的な抗菌性物質を合成しようとしても、どのような化学構造としたらよいのかが分からないという問題がある。また、たとえ人工的な抗菌性物質を見出したとしても、複雑な合成プロセスが必要であって技術的な困難性を有するのでは、製造コストが高騰化するという問題がある。さらには、ペプチドからなる抗菌剤ではアミノ酸配列を変化させることで二次構造も変化してしまう可能性が高く、その場合、細菌の細胞膜との相互作用に影響を生じ、抗菌性を消失すると考えられる。こうした人工的な抗菌性物質の合成の困難性から、抗菌剤は天然物から得られた抗菌物質と類似の化合物を合成し、その抗菌活性を調べることが主流とされていた。 In many cases, the mechanism of antibacterial action has not been elucidated in detail, and there is a problem that even if an attempt is made to synthesize an artificial antibacterial substance, it is not known what chemical structure it should be. Moreover, even if an artificial antibacterial substance is found, there is a problem that the manufacturing cost increases if a complicated synthesis process is required and technical difficulties are involved. Furthermore, antibacterial agents consisting of peptides are likely to change the secondary structure by changing the amino acid sequence. In this case, the interaction with the bacterial cell membrane is affected, and the antibacterial properties are lost. It is done. Because of the difficulty in synthesizing artificial antibacterial substances, antibacterial agents mainly synthesize compounds similar to antibacterial substances obtained from natural products and investigate their antibacterial activity.
本発明は、上記従来の実情に鑑みてなされたものであって、抗菌性に優れ、少ないプロセスのために合成が容易で安価に大量合成が可能な化合物、その化合物の製造方法およびその化合物を用いた抗菌剤を提供することを解決すべき課題としている。また、その抗菌剤を合成するための試薬を提供することも解決すべき課題としている。 The present invention has been made in view of the above-described conventional circumstances, and has excellent antibacterial properties, is easy to synthesize for a small number of processes, can be synthesized in large quantities at low cost, a method for producing the compound, and a compound thereof. Providing the used antibacterial agent is a problem to be solved. In addition, providing a reagent for synthesizing the antibacterial agent is also a problem to be solved.
まず本発明者は、以下に示すシクロデキストリンの化学構造に着目し、これが新たな抗菌物質に利用できるのではないかと考え、次のような分子設計を考えた。 First, the present inventor focused on the following chemical structure of cyclodextrin, thought that it could be used as a new antibacterial substance, and considered the following molecular design.
シクロデキストリンは単糖であるグルコースが環状に連なった約１ナノメータの直径を持つ環状糖質である。グルコースの数が８個のものは、γシクロデキストリン[1]と呼ばれている（下記化１参照）。シクロデキストリンのグルコースの６位に存在する第一級水酸基は環状構造の一方の開口部に、２位と３位に存在する第二級水酸基は他方の開口部に配向している。また、上記それぞれの水酸基は反応性に違いがあり、選択的に化学修飾をすることが可能である。 Cyclodextrin is a cyclic carbohydrate having a diameter of about 1 nanometer in which glucose, which is a monosaccharide, is linked in a ring. Those having 8 glucoses are called γ-cyclodextrins [1] (see Chemical Formula 1 below). The primary hydroxyl group present at the 6-position of glucose in the cyclodextrin is oriented at one opening of the cyclic structure, and the secondary hydroxyl groups present at the 2- and 3-positions are oriented at the other opening. Further, each of the above hydroxyl groups has a difference in reactivity, and can be selectively chemically modified.
環状糖質であるシクロデキストリンには、それを構成するグルコースの数が異なる類縁体があり、グルコースの数が５個以上のものが知られている。また、グルコースは２つ以上が鎖状に連なり、オリゴ糖〜多糖を形成する。これにはアミロースに代表されるグルカンが含まれる。さらに、グルコースの異性体である単糖が連なった多糖があり、マンノースを繰り返し構造とするマンナンなどがある。これらもまた同様に、その繰り返し構造である単糖の上に反応性の異なる水酸基を持つため、選択的に化学修飾をすることが可能である。 Cyclodextrins, which are cyclic carbohydrates, have analogs with different numbers of glucose, and those with 5 or more glucoses are known. In addition, two or more glucoses are linked in a chain to form oligosaccharides to polysaccharides. This includes glucans represented by amylose. Furthermore, there are polysaccharides in which monosaccharides that are isomers of glucose are linked, and there are mannans having mannose as a repeating structure. Similarly, these have hydroxyl groups with different reactivities on the monosaccharide as the repeating structure, and therefore can be selectively chemically modified.
細菌に抗菌性を示すペプチドであるポリミキシンＢの構造を化２に示す。これは天然に存在する環状ペプチドである。ポリミキシンＢは、グラム陰性菌の外膜中のリポ多糖と結合して膜構造を破壊する。さらに細胞質膜をも傷害して抗菌性を示す。それを可能にする要因である構造的な特徴として、陽イオン性アミノ基と疎水性基をそれぞれ複数有することが挙げられる。 The structure of polymyxin B, a peptide that exhibits antibacterial properties against bacteria, is shown in Chemical Formula 2. This is a naturally occurring cyclic peptide. Polymyxin B binds to lipopolysaccharide in the outer membrane of Gram-negative bacteria and destroys the membrane structure. Furthermore, it damages the cytoplasmic membrane and exhibits antibacterial properties. A structural feature which is a factor that makes it possible is to have a plurality of cationic amino groups and hydrophobic groups.
そこで、本発明者は細菌への抗菌性を実現するべく、シクロデキストリン上に陽イオン性アミノ基と疎水性アルキル基を複数個集積させることを考えた。このアミノ基がまず菌体膜のリン酸部と結合し、引き続きアルキル鎖が膜内部に侵入することで膜構造を破壊すると予想したのである。 Therefore, the present inventor considered to accumulate a plurality of cationic amino groups and hydrophobic alkyl groups on cyclodextrin in order to realize antibacterial properties against bacteria. It was predicted that this amino group would first bind to the phosphoric acid part of the cell membrane, and then the alkyl chain would break into the membrane, thereby destroying the membrane structure.
アミノ基の形成については、シクロデキストリン上に導入したアジド基を化学変換することでアミノ基を導入する方法を検討した。この化学変換の鍵段階はクリックケミストリーを採用した。クリックケミストリーとは、以下のような特徴を有する反応のことである。
（１）汎用性が高い
（２）高効率、高収率
（３）副生成物を生じない
（４）立体特異的である
（５）精製が容易である
（６）出発物質と試薬が手に入れやすい。
Regarding the formation of the amino group, a method for introducing an amino group by chemically converting the azide group introduced onto the cyclodextrin was examined. The key step of this chemical transformation was click chemistry. Click chemistry is a reaction having the following characteristics.
(1) High versatility (2) High efficiency, high yield (3) No by-product (4) Stereospecific (5) Easy purification (6) Handy starting materials and reagents Easy to put in.
クリックケミストリーとして最も一般的なものとされているのがＨｕｉｓｇｅｎ反応である（化３参照）。この反応は、アジドと末端アルキンがトリアゾール環を形成する反応である。銅を触媒にすることで１，４-二置換体が選択的に形成し、かつ反応は大幅に加速する。また、アジドとアルキンは互いの間でのみ穏やかに反応し、そして標準的な反応条件でよく用いられる求核剤、求電子剤および溶媒に対して安定である。生成するトリアゾール環も安定な官能基である。このような特徴からＨｕｉｓｇｅｎ反応は広い分野で利用されている。 The most common click chemistry is the Huisgen reaction (see Chemical Formula 3). This reaction is a reaction in which an azide and a terminal alkyne form a triazole ring. By using copper as a catalyst, a 1,4-disubstituted product is selectively formed, and the reaction is greatly accelerated. Also, azides and alkynes react only mildly between each other and are stable to nucleophiles, electrophiles and solvents often used in standard reaction conditions. The resulting triazole ring is also a stable functional group. Due to these characteristics, the Huisgen reaction is used in a wide range of fields.
本発明者らは、このＨｕｉｓｇｅｎ反応を利用し、シクロデキストリンに1級アミノ基を導入した化合物が抗菌剤として機能することを見出し、既に特許出願を行っている（特願２０１２−４０９３４）。 The present inventors have found that a compound in which a primary amino group is introduced into cyclodextrin functions as an antibacterial agent using this Huisgen reaction, and has already filed a patent application (Japanese Patent Application No. 2012-40934).
さらに、発明者らは、Ｈｕｉｓｇｅｎ反応を利用し、シクロデキストリンに２級アミノ基を導入した化合物の合成にも成功し、この化合物が抗菌剤として機能することを見出し、本発明を完成するに至った。
ここではシクロデキストリンから少ないプロセスで抗菌剤が容易に合成でき、数十段階以上のプロセスが必要な抗菌ペプチド合成に比べて著しく優れている。
Furthermore, the inventors succeeded in synthesizing a compound in which a secondary amino group is introduced into cyclodextrin by utilizing the Huisgen reaction, and found that this compound functions as an antibacterial agent, and completed the present invention. It was.
Here, an antibacterial agent can be easily synthesized from cyclodextrin with a small number of processes, and it is remarkably superior to the synthesis of antibacterial peptides that require several tens of steps or more.
すなわち、本発明は下記一般式で示される、単糖がグリコシド結合で鎖状又は環状に連結した糖誘導体又はその塩である。ここで、トリアゾールの窒素は単糖の一級水酸基と置換している。さらに、R１はアルキレン基を示し、R２は分岐してもよくシクロ環を有してもよいアルキル基を示す。ｎは２以上の整数を示す。 That is, the present invention is a sugar derivative or a salt thereof, represented by the following general formula, in which monosaccharides are linked in a chain or cyclic manner by glycosidic bonds. Here, the nitrogen of the triazole is substituted with a primary hydroxyl group of a monosaccharide. Further, R 1 represents an alkylene group, and R 2 represents an alkyl group that may be branched or have a cyclo ring. n represents an integer of 2 or more.
ここで、R1はメチレン基とすることが好ましい。 Here, R 1 is preferably a methylene group.
R2は炭素数７個、６個、５個又は４個からなるアルキル基であることが好ましい。
また、R2は窒素に結合するメチレン基を有することが好ましい。
R 2 is preferably an alkyl group having 7, 6, 5 or 4 carbon atoms.
R 2 preferably has a methylene group bonded to nitrogen.
R2は２−メチルプロピル基、２−エチルブチル基、シクロブチルメチル基、シクロペンチルメチル基、又はシクロへシルメチル基であることが好ましい。 R 2 is preferably a 2-methylpropyl group, a 2-ethylbutyl group, a cyclobutylmethyl group, a cyclopentylmethyl group, or a cyclohexylmethyl group.
ｎは実際に存在するオリゴ糖又は多糖を構成する数とする。例えば、１０００以下とすることができる。ｎは５〜５０とすることが好ましい。 n is a number that constitutes an actually existing oligosaccharide or polysaccharide. For example, it can be 1000 or less. n is preferably 5 to 50.
本発明の化合物としては、単糖がグルコース、マンノース、フルクトース、またはガラクトースに代表される炭素６個からなるヘキソースである糖誘導体又はその塩が挙げられる。 Examples of the compound of the present invention include a sugar derivative or a salt thereof in which a monosaccharide is a hexose having 6 carbons represented by glucose, mannose, fructose, or galactose.
また、本発明の化合物としては、単糖がグルコースであり、このグルコースが環状に連結したオリゴ糖誘導体又はその塩が挙げられる。 In addition, examples of the compound of the present invention include an oligosaccharide derivative in which the monosaccharide is glucose and the glucose is linked in a cyclic manner, or a salt thereof.
より具体的には、ｎが８であり、グルコースが環状に連結したオリゴ糖の誘導体又はその塩が挙げられる。 More specifically, a derivative of oligosaccharide or a salt thereof in which n is 8 and glucose is linked in a cyclic manner.
また、本発明の抗菌剤は、上記した糖誘導体又はその塩を有効成分として含有することを特徴とする。ここで「有効成分として含有する」には、糖誘導体又はその塩が抗菌剤の構造全体を構成する場合に限らず、抗菌剤の構造の一部を構成する場合も含まれる。 The antibacterial agent of the present invention is characterized by containing the above-described sugar derivative or salt thereof as an active ingredient. Here, “containing as an active ingredient” includes not only the case where the sugar derivative or a salt thereof constitutes the entire structure of the antibacterial agent, but also the case where it constitutes a part of the structure of the antibacterial agent.
また、本発明は、抗菌剤を合成するための試薬としての、下記一般式で示される末端アルキンである。ただし、R１はアルキレン基、R２は分岐してもよくシクロ環を有してもよいアルキル基を示し、R3は水素又は脱保護可能な置換基を示す。この置換基は、例えばカルボニル基によってＮに結合した基を示す。 Moreover, this invention is the terminal alkyne shown by the following general formula as a reagent for synthesize | combining an antibacterial agent. However, R 1 represents an alkylene group, R 2 represents an alkyl group which may be branched or have a cyclo ring, and R 3 represents hydrogen or a deprotectable substituent. This substituent is, for example, a group bonded to N by a carbonyl group.
ここで、R1はメチレン基とすることが好ましい。 Here, R 1 is preferably a methylene group.
R2は炭素数７個、６個、５個又は４個からなるアルキル基であることを特徴とする。 R 2 is characterized by being an alkyl group comprising 7, 6, 5 or 4 carbon atoms.
また、R2は窒素に結合するメチレン基を有することが好ましい。 R 2 preferably has a methylene group bonded to nitrogen.
R2は２−メチルプロピル基、２−エチルブチル基、シクロブチルメチル基、シクロペンチルメチル基、又はシクロへシルメチル基であることが好ましい。 R 2 is preferably a 2-methylpropyl group, a 2-ethylbutyl group, a cyclobutylmethyl group, a cyclopentylmethyl group, or a cyclohexylmethyl group.
R3は第三ブトキシカルボニル基であることが好ましい。 R 3 is preferably a tertiary butoxycarbonyl group.
また、本発明は、抗菌剤を合成するための試薬として、下記一般式で示される、単糖がグリコシド結合で鎖状または環状に連結した糖のアジ化物である。ここで、アジド基は単糖の一級水酸基と置換している。ｎは２以上の整数を示す。 The present invention is also a reagent for synthesizing an antibacterial agent, which is an azide of a sugar represented by the following general formula, in which monosaccharides are linked in a chain or cyclic manner with glycosidic bonds. Here, the azide group is substituted with a primary hydroxyl group of a monosaccharide. n represents an integer of 2 or more.
ｎは実際に存在するオリゴ糖〜多糖を構成する数とする。例えば、１０００以下とすることができる。ｎは５〜５０とすることが好ましい。 n is a number that constitutes an actually existing oligosaccharide to polysaccharide. For example, it can be 1000 or less. n is preferably 5 to 50.
本発明の糖のアジ化物としては、単糖がグルコース、マンノース、フルクトース、またはガラクトース、に代表される炭素６個からなるヘキソースであるオリゴ糖又は多糖のアジ化物が挙げられる。 Examples of the sugar azide of the present invention include an azide of an oligosaccharide or polysaccharide whose monosaccharide is a hexose consisting of 6 carbons represented by glucose, mannose, fructose, or galactose.
また、本発明の糖のアジ化物としては、単糖グルコースが環状に連結したオリゴ糖のアジ化物が挙げられる。 The azide of the sugar of the present invention includes an azide of an oligosaccharide in which monosaccharide glucose is linked in a cyclic manner.
より具体的には、ｎが８であり、グルコースが環状に連結したオリゴ糖のアジ化物が挙げられる。 More specifically, an azide of an oligosaccharide in which n is 8 and glucose is linked in a cyclic manner can be mentioned.
下記一般式で示される、単糖がグリコシド結合で鎖状または環状に連結したオリゴ糖又は多糖のアジ化物と、下記一般式で示される末端アルキンとを反応させることを特徴とする請求項１〜８に記載の糖誘導体又はその塩の製造方法。なお、この製造方法においては、上記反応後に、必要に応じて、脱保護を行ってもよい。 An azide of an oligosaccharide or polysaccharide in which monosaccharides represented by the following general formula are linked in a chain or cyclic form with glycosidic bonds and a terminal alkyne represented by the following general formula are reacted: 9. A process for producing the sugar derivative or salt thereof according to 8. In this production method, deprotection may be performed as necessary after the above reaction.
(一般式中のアジド基は単糖の一級水酸基と置換している。また、ｎについては、前述の糖のアジ化物と同じである。) (The azide group in the general formula is substituted with the primary hydroxyl group of a monosaccharide. Also, n is the same as the above-mentioned azide of sugar.)
（一般式中のＲ１、Ｒ２、Ｒ３については、前述の末端アルキンと同じである。） (R1, R2, and R3 in the general formula are the same as the above-mentioned terminal alkyne.)
＜分子設計＞
本発明の抗菌物質を合成するにあたっては、以下のコンセプトにより分子設計を行った。
細菌膜を傷害して細菌への抗菌性を実現するためには、親水性基と疎水性基が複数個存在し、かつそのバランスが重要になる。そこで、シクロデキストリン上にアルキルアミノメチルトリアゾール基を集積させる。そのために親水性であるシクロデキストリンのグルコース部分にトリアゾール部を介してアミノメチル部を連結することで、親水性でかつ陽イオン性部分を構成する。さらにアミノ基に結合したアルキル基が疎水性部分を形成する。細菌膜を傷害する際には、このアミノ基がまず細菌膜の陰イオン性リン酸部と結合し、引き続き疎水性アルキル部が膜内部に侵入することで膜構造を破壊する。
このようにアルキルアミノアルキルトリアゾール基を導入したグルコースを複数個含むことが、シクロデキストリン誘導体が抗菌物質として機能するために重要である。したがって同様にグルコースが鎖状に連なって形成された、オリゴ糖又は多糖グルカンにおいて、そのグルコース部に同様にアルキルアミノアルキルトリアゾール基を導入すれば抗菌性を現すと考えられる。
さらに、グルコースの異性体にはマンノースに代表される単糖ヘキソースがある。これらはグルコースと同じ数の炭素と水酸基を持ち、同様の親水性を持つ。そして、それが連なったオリゴ糖〜多糖が存在し、代表としてマンナンがある。これらもまた、その繰り返し構造である単糖の上にアルキルアミノアルキルトリアゾール基を導入すれば抗菌性を現すと考えられる。
<Molecular design>
In synthesizing the antibacterial substance of the present invention, molecular design was performed based on the following concept.
In order to injure the bacterial membrane and realize antibacterial properties against bacteria, there are a plurality of hydrophilic groups and hydrophobic groups, and the balance is important. Therefore, alkylaminomethyltriazole groups are accumulated on the cyclodextrin. Therefore, a hydrophilic and cationic part is constituted by connecting an aminomethyl part via a triazole part to a glucose part of cyclodextrin which is hydrophilic. Furthermore, the alkyl group bonded to the amino group forms a hydrophobic portion. When damaging the bacterial membrane, the amino group first binds to the anionic phosphate portion of the bacterial membrane, and subsequently the hydrophobic alkyl portion penetrates into the membrane to destroy the membrane structure.
Thus, it is important for the cyclodextrin derivative to function as an antibacterial substance to contain a plurality of glucoses into which an alkylaminoalkyltriazole group has been introduced. Therefore, in an oligosaccharide or polysaccharide glucan in which glucose is similarly formed in a chain, it is considered that antibacterial properties are exhibited if an alkylaminoalkyltriazole group is similarly introduced into the glucose portion.
Furthermore, glucose isomers include monosaccharide hexoses represented by mannose. These have the same number of carbon and hydroxyl groups as glucose and have similar hydrophilicity. There are oligosaccharides to polysaccharides in which they are connected, and mannan is a representative example. These are also considered to exhibit antibacterial properties when an alkylaminoalkyltriazole group is introduced onto a monosaccharide having a repeating structure.
そして、上記のいずれか１項に記載のオリゴ糖〜多糖の誘導体又はその塩を有効成分として含有するものは抗菌剤となる。ここで「有効成分として含有する」には、オリゴ糖〜多糖の誘導体又はその塩が抗菌剤の構造全体を構成する場合に限らず、抗菌剤の構造の一部を構成する場合も含まれる。 And what contains the derivative | guide_body or its salt of the oligosaccharide-polysaccharide of any one of said 1 as an active ingredient becomes an antibacterial agent. Here, “containing as an active ingredient” includes not only the case where the oligosaccharide to polysaccharide derivative or salt thereof constitutes the entire structure of the antibacterial agent, but also the case where it constitutes a part of the structure of the antibacterial agent.
上記のアルキルアミノメチルトリアゾール基の導入については、シクロデキストリン上に形成したアジド基を化学変換することで導入する方法を検討した。この化学変換法としては、クリック反応として最も一般的なＨｕｉｓｇｅｎ反応を採用した（前述した化３に示す反応式を参照されたい）。この反応は、アジドと末端アルキンがトリアゾール環を形成する反応である。銅を触媒にすることで１，４-二置換体が選択的に形成し、かつ反応は大幅に加速する。また、アジドとアルキンは互いの間でのみ穏やかに反応し、そして標準的な反応条件でよく用いられる求核剤、求電子剤および溶媒に対して安定である。 Regarding the introduction of the above alkylaminomethyltriazole group, a method of introducing the azide group formed on the cyclodextrin by chemical conversion was examined. As this chemical conversion method, the most common Huisgen reaction was adopted as the click reaction (see the reaction formula shown in Chemical Formula 3 above). This reaction is a reaction in which an azide and a terminal alkyne form a triazole ring. By using copper as a catalyst, a 1,4-disubstituted product is selectively formed, and the reaction is greatly accelerated. Also, azides and alkynes react only mildly between each other and are stable to nucleophiles, electrophiles and solvents often used in standard reaction conditions.
以下に示す実施例１〜５のシクロデキストリン誘導体（ａ）〜（ｅ）を合成した。以下、合成法について詳細に記載する。 The following cyclodextrin derivatives (a) to (e) of Examples 1 to 5 were synthesized. Hereinafter, the synthesis method will be described in detail.
（オクタアジ化γシクロデキストリン[３]の合成）
下記化６に示すように、Ｈｕｉｓｇｅｎ反応の原料となるオクタアジ化されたγシクロデキストリン[３]を、オクタクロロ化シクロデキストリン[２]を経由して合成した。具体的には、シクロデキストリン[１]をジメチルホルムアミドに溶解し、これに塩化メタンスルホニルを加えて６５℃にて２０時間攪拌することでオクタクロロ化シクロデキストリン[２]を得た。これをジメチルホルムアミドに溶解した後、アジ化ナトリウムを加え、１００℃にて１７時間反応してオクタアジ化γシクロデキストリン[３]を得た。その構造はＨ−ＮＭＲ、ＩＲによって確認した。
(Synthesis of octaazidated γ cyclodextrin [3])
As shown in the following chemical formula 6, octaazidized γ-cyclodextrin [3], which is a raw material for the Huisgen reaction, was synthesized via octachlorinated cyclodextrin [2]. Specifically, cyclodextrin [1] was dissolved in dimethylformamide, methanesulfonyl chloride was added thereto, and the mixture was stirred at 65 ° C. for 20 hours to obtain octachlorinated cyclodextrin [2]. This was dissolved in dimethylformamide, sodium azide was added, and the mixture was reacted at 100 ° C. for 17 hours to obtain octaazidized γ cyclodextrin [3]. The structure was confirmed by H-NMR and IR.
以下、オクタクロロ化シクロデキストリン[２]及びオクタアジ化γシクロデキストリン[３]の合成方法及び化合物データについて具体的に述べる。
（オクタクロロ化シクロデキストリン[２]）
凍結乾燥したγシクロデキストリン[1] 2.11 g (1.62 mmol) を乾燥DMF 20 mlに溶解し、氷浴中で塩化メタンスルホニル 7.44 g (64.9 mmol) を加えた。アルゴン雰囲気下、室温で30分間攪拌し、その後、65 ℃の油浴中で20時間攪拌した。溶媒を減圧留去し、残渣にメタノール10 mlを加えた。氷浴中で3 M ナトリウムメトキシド/メタノール溶液をpH 9になるまで加え、生じた沈殿をろ取した。これを氷水を用いて洗浄し、凍結乾燥し、白色固体の目的物1.94 g (1.34 mmol, 83%) を得た。
Rf = 0.62 (H2O/1-PrOH/AcOEt=5/7/7 (v/v/v))
1H NMR (400 MHz, DMSO-d6) δ＝3.33-3.40 (H-2,4), 3.60 (H-3), 3.82-3.84 (H-5,6), 4.02 (H-6), 4.98 (H-1), 5.94 (OH-3), 5.98 (OH-2)
MALDI-TOF-MS: [M+Na]+ Calcd. for C48H72 35Cl6 37Cl2NaO32: 1467, Found: 1467.
[M+K]+ Calcd. for C48H72 35Cl6 37Cl2KO32: 1483, Found: 1483.
（オクタアジ化γシクロデキストリン[３]）
オクタクロロ化シクロデキストリン [２] 1.00 g (692 μmol) を乾燥DMF 20 mlに溶解し、アジ化ナトリウム1.80 g (27.7 mmol) を加え、アルゴン雰囲気下、100 ℃の油浴中で17時間攪拌した。溶媒を減圧留去し、水を用いた洗浄を3回行い、得られた固体を凍結乾燥し、白色固体の目的物909 mg (607 μmol, 88%) を得た。
Rf = 0.62 (H2O/1-PrOH/AcOEt=5/7/7 (v/v/v))
1H NMR (400 MHz, DMSO-d6):δ＝3.32-3.40 (H-4,6), 3.54-3.60 (H-5,6), 3.71-3.74 (H-2,3), 4.93 (H-1), 5.88-5.94 (OH)
FT-IR (KBr, cm-1) 669.18, 757.89, 842.74, 943.02, 1049.09, 1078.98, 1156.12, 1288.22, 1437.67, 1630.52, 1741.41, 2105.89, 2925.48, 3397.96
（実施例１）
・化合物（ａ）の合成
上記のようにして合成したオクタアジ化γシクロデキストリン[３] 200 mg (134 mmol) をDMSO 10 ml に溶解し、アスコルビン酸ナトリウム水溶液(265 mg/ml) を1 ml (265 mg, 1.34 mmol)、硫酸銅水溶液 (26.8 mg/ml) を1 ml (26.8 mg, 107 mmol)、シクロヘキシルメチルプロパルギルアミン(250 mg, 1.65 mmol)のDMSO溶液10 ml を加え、マイクロ波加熱（120 ℃）下で10分間攪拌した。これをろ過した後、溶媒を減圧留去し、残渣をアセトンで洗浄し、引き続き10% アンモニア水で洗浄し、得られた固体を凍結乾燥し、シクロヘキシルメチルアミノ部を持つ実施例１の化合物（ａ） 312 mg (86%) を得た
Rf = 0.00 (H2O/1-propanol/AcOEt (5/7/7))
1H NMR (400 MHz, DMSO-d6): d = 0.78-1.59 (cyclohexane), 2.27-2.33 (CH2), 3.35-4.20 ( H-2,3,4,5,6), 5.09 (H-1), 6.02-6.17 (OH), 7.73 (triazole)
13C NMR (125 MHz, DMSO-d6): d = 25.6, 26.3, 31.0 (CH2), 37.3 (CH), 44.2 (CCH2NH), 49.5 (C-6), 55.5 (CH2NH), 69.8 (C-5), 72.1, 72.3 (C-2,3), 82.4 (C-4), 101.6 (C-1), 123.4 (C-5 triazole), 146.0 (C-4 triazole)
MALDI-TOF-MS: [M + K]+ Calcd. for C128H208KN32O32: 2744.5, Found: 2744.7.
Anal.: + 7.5H2O Calcd. for C128H223N32O39.5: C 54.09, H 7.91, N 15.77. Found: C 54.47, H 7.35, N 15.04.
FT-IR: (KBr, cm-1) 598.8, 1044.3, 1079.0, 1151.2, 1607.4, 1644.0, 2850.3, 2923.6, 3379.6
（実施例２）
シクロデキストリン誘導体（ｂ）の合成
（アセチル化オクタアジ化γシクロデキストリン[４]の合成）
Hereafter, the synthesis method and compound data of octachloroated cyclodextrin [2] and octaazidated γ cyclodextrin [3] will be specifically described.
(Octachlorinated cyclodextrin [2])
Lyophilized γ-cyclodextrin [1] 2.11 g (1.62 mmol) was dissolved in 20 ml of dry DMF, and 7.44 g (64.9 mmol) of methanesulfonyl chloride was added in an ice bath. The mixture was stirred at room temperature for 30 minutes under an argon atmosphere, and then stirred in an oil bath at 65 ° C. for 20 hours. The solvent was distilled off under reduced pressure, and 10 ml of methanol was added to the residue. A 3 M sodium methoxide / methanol solution was added in an ice bath until pH 9 was reached, and the resulting precipitate was collected by filtration. This was washed with ice water and freeze-dried to obtain 1.94 g (1.34 mmol, 83%) of the desired product as a white solid.
R f = 0.62 (H 2 O / 1-PrOH / AcOEt = 5/7/7 (v / v / v))
1 H NMR (400 MHz, DMSO-d 6 ) δ = 3.33-3.40 (H-2,4), 3.60 (H-3), 3.82-3.84 (H-5,6), 4.02 (H-6), 4.98 (H-1), 5.94 (OH-3), 5.98 (OH-2)
MALDI-TOF-MS: [M + Na] + Calcd. For C 48 H 72 35 Cl 6 37 Cl 2 NaO 32 : 1467, Found: 1467.
[M + K] + Calcd. For C 48 H 72 35 Cl 6 37 Cl 2 KO 32 : 1483, Found: 1483.
(Octazidated gamma cyclodextrin [3])
Octachloroated cyclodextrin [2] 1.00 g (692 μmol) was dissolved in 20 ml of dry DMF, 1.80 g (27.7 mmol) of sodium azide was added, and the mixture was stirred in an oil bath at 100 ° C. for 17 hours under an argon atmosphere. The solvent was distilled off under reduced pressure, followed by washing with water three times. The obtained solid was lyophilized to obtain 909 mg (607 μmol, 88%) of the desired product as a white solid.
R f = 0.62 (H 2 O / 1-PrOH / AcOEt = 5/7/7 (v / v / v))
1 H NMR (400 MHz, DMSO-d 6 ): δ = 3.32-3.40 (H-4,6), 3.54-3.60 (H-5,6), 3.71-3.74 (H-2,3), 4.93 ( H-1), 5.88-5.94 (OH)
FT-IR (KBr, cm -1 ) 669.18, 757.89, 842.74, 943.02, 1049.09, 1078.98, 1156.12, 1288.22, 1437.67, 1630.52, 1741.41, 2105.89, 2925.48, 3397.96
Example 1
Synthesis of Compound (a) 200 mg (134 mmol) of octaazidated γ cyclodextrin [3] synthesized as described above was dissolved in 10 ml of DMSO, and 1 ml of sodium ascorbate aqueous solution (265 mg / ml) ( 265 mg, 1.34 mmol), 1 ml (26.8 mg, 107 mmol) of an aqueous copper sulfate solution (26.8 mg / ml), 10 ml of DMSO solution of cyclohexylmethylpropargylamine (250 mg, 1.65 mmol), and microwave heating ( (120 ° C.) for 10 minutes. After filtration, the solvent was distilled off under reduced pressure, the residue was washed with acetone and subsequently with 10% aqueous ammonia, and the resulting solid was lyophilized to give the compound of Example 1 having a cyclohexylmethylamino moiety ( a) 312 mg (86%) was obtained
R f = 0.00 (H 2 O / 1-propanol / AcOEt (5/7/7))
1 H NMR (400 MHz, DMSO-d 6 ): d = 0.78-1.59 (cyclohexane), 2.27-2.33 (CH 2 ), 3.35-4.20 (H-2,3,4,5,6), 5.09 (H -1), 6.02-6.17 (OH), 7.73 (triazole)
13 C NMR (125 MHz, DMSO-d 6 ): d = 25.6, 26.3, 31.0 (CH 2 ), 37.3 (CH), 44.2 (CCH 2 NH), 49.5 (C-6), 55.5 (CH 2 NH) , 69.8 (C-5), 72.1, 72.3 (C-2,3), 82.4 (C-4), 101.6 (C-1), 123.4 (C-5 triazole), 146.0 (C-4 triazole)
MALDI-TOF-MS: [M + K] + Calcd. For C 128 H 208 KN 32 O 32 : 2744.5, Found: 2744.7.
Anal .: + 7.5H 2 O Calcd. For C 128 H 223 N 32 O 39.5 : C 54.09, H 7.91, N 15.77. Found: C 54.47, H 7.35, N 15.04.
FT-IR: (KBr, cm -1 ) 598.8, 1044.3, 1079.0, 1151.2, 1607.4, 1644.0, 2850.3, 2923.6, 3379.6
(Example 2)
Synthesis of cyclodextrin derivative (b) (synthesis of acetylated octaazidized γ cyclodextrin [4])
オクタアジ化γシクロデキストリン [３] 108 mg (72.3 μmol) を乾燥ピリジン3 mlで3回共沸し、そこに乾燥ピリジン1 ml、無水酢酸1 ml (10.6 mmol) を加え、アルゴン雰囲気下、室温で15時間攪拌した。溶液を減圧留去し、残渣にジクロロメタン20 mlを加え、水を用いた洗浄、飽和食塩水を用いた脱水後、硫酸ナトリウムを用いて乾燥した。それを順相シリカゲルカラム (ヘキサン/酢酸エチル ) にて精製した。該当分画を回収し, 溶媒を減圧留去し, 白色結晶[４]121 mg (56.0 μmol, 77%) を得た。
Rf = 0.58 (CH2Cl2/CH3OH=9/1 (v/v))
1H NMR (400 MHz, CDCl3): δ＝2.08-2.09 (CH3), 3.60 (H-6), 3.71-3.79 (H-4,6), 3.96-3.98 (H-5), 4.77 (H-2), 5.16 (H-1), 5.32 (H-3)
MALDI-TOF-MS: [M + Na]+ Calcd. for C80H104N24NaO48: 2192, Found: 2192.
[M + K]+ Calcd. for C80H104KN24O48: 2208, Found: 2208.
(アセチル化オクタアジ化γシクロデキストリン[４]を用いたクリック反応)
硫酸銅 9.29 mg(3.69×10-5 mol)を120 μlの純水に溶かし、アスコルビン酸ナトリウム91.5 mg(4.61×10-4 mol)を280 μlの純水に溶かしたものを加えた。その溶液を、DMSO 2 mlに溶かした化合物[４] 100 mg(4.61×10-5 mol)に加えた。ここにDMSO2.5 mlに溶かした第三ブトキシカルボニル化２−エチルブチルアミノプロピン110.3 mg(4.61×10-4 mol)を加えた。これにマイクロ波を照射し加熱(10 min, 120℃)した後、酢酸エチルに溶かし、これを5 %EDTA二ナトリウム水溶液で洗浄し、減圧留去にて溶媒を除去した残渣をシリカゲルカラム(ヘキサン/酢酸エチル) により精製し、白色固体143 mgを得た(収率75.9 %)。
TLC Rf＝0.74 (ヘキサン/酢酸エチル=1/9)
1H NMR (400 MHz, CDCl3):
δ= 0.82-0.86 (CH3), 1.24-1.28 (CH2), 1.41 (C(CH3)3), 1.60 (CH), 2.04-2.05 (COCH3), 3.11-3.16(CH2), 3.58 (H-4), 4.35-4.49 (H-5, 6), 4.67-4.69 (H-2), 4.83-4.98 (CH2), 5.39-5.40 (H-3), 5.65 (H-1), 7.71 (triazole)
13C NMR (100 MHz, CDCl3): d = 10.6-10.6 (CH3), 20.7-20.8 (CH3-acetyl), 23.1 (CH2), 28.0-28.4 ((CH3)3-Boc), 29.7 (CH), 39.0-39.4 (CH), 42.4 (C6), 49.8 (CH2-triazole), 50.4 (CH2-N(Boc)), 69.7-70.8 (C5, C2, C3, C4), 79.7 (C(CH3)3-Boc), 95.6 (C1), 124.7-125.3 (CH-triazole), 145.3 (=C-N-triazole), 155.5-155.9 (C=O-Boc) , 169.69-170.2 (C=O-acetyl)
FT-IR :(KBr, cm-1) 782.958, 878.417, 949.77, 1045.23, 1168.65, 1242.9, 1367.28, 1416.46, 1463.71, 1689.34, 1758.76, 2877.27, 2934.16, 2965.02, 3434.6
Calcd. for C192H304N32O64+ 2.0 H2O: C 55.96, H 7.53, N 10.88. Found: C 55.95, H 7.58, N 10.64
(第三ブトキシカルボニル基の脱保護)
上記のクリック反応生成物18.4 mg(4.50×10-6 mol)にトリフルオロ酢酸500 mlを加えて溶解させた。その後、２０分攪拌し、減圧留去により白色固体18.9 mg (収率100 %)を得た。
Octazylated γ-cyclodextrin [3] 108 mg (72.3 μmol) was azeotroped 3 times with 3 ml of dry pyridine, and 1 ml of dry pyridine and 1 ml (10.6 mmol) of acetic anhydride were added thereto, and at room temperature under argon atmosphere. Stir for 15 hours. The solution was evaporated under reduced pressure, 20 ml of dichloromethane was added to the residue, washed with water, dehydrated with saturated brine, and dried over sodium sulfate. It was purified on a normal phase silica gel column (hexane / ethyl acetate). The relevant fractions were collected and the solvent was distilled off under reduced pressure to obtain 121 mg (56.0 μmol, 77%) of white crystals [4].
R f = 0.58 (CH 2 Cl 2 / CH 3 OH = 9/1 (v / v))
1 H NMR (400 MHz, CDCl 3 ): δ = 2.08-2.09 (CH 3 ), 3.60 (H-6), 3.71-3.79 (H-4,6), 3.96-3.98 (H-5), 4.77 ( H-2), 5.16 (H-1), 5.32 (H-3)
MALDI-TOF-MS: [M + Na] + Calcd. For C 80 H 104 N 24 NaO 48 : 2192, Found: 2192.
[M + K] + Calcd. For C 80 H 104 KN 24 O 48 : 2208, Found: 2208.
(Click reaction using acetylated octaazidized γ cyclodextrin [4])
Copper sulfate 9.29 mg (3.69 × 10 −5 mol) dissolved in 120 μl of pure water and sodium ascorbate 91.5 mg (4.61 × 10 −4 mol) dissolved in 280 μl of pure water were added. The solution was added to 100 mg (4.61 × 10 −5 mol) of the compound [4] dissolved in 2 ml of DMSO. To this was added 110.3 mg (4.61 × 10 −4 mol) of tert-butoxycarbonylated 2-ethylbutylaminopropyne dissolved in 2.5 ml of DMSO. This was irradiated with microwaves and heated (10 min, 120 ° C), then dissolved in ethyl acetate, washed with 5% aqueous EDTA disodium solution, and the solvent was removed by distillation under reduced pressure. / Ethyl acetate) to obtain 143 mg of a white solid (yield 75.9%).
TLC R f = 0.74 (hexane / ethyl acetate = 1/9)
1 H NMR (400 MHz, CDCl 3 ):
δ = 0.82-0.86 (CH 3 ), 1.24-1.28 (CH 2 ), 1.41 (C (CH 3 ) 3 ), 1.60 (CH), 2.04-2.05 (COCH 3 ), 3.11-3.16 (CH 2 ), 3.58 (H-4), 4.35-4.49 (H-5, 6), 4.67-4.69 (H-2), 4.83-4.98 (CH 2 ), 5.39-5.40 (H-3), 5.65 (H-1), 7.71 (triazole)
13 C NMR (100 MHz, CDCl 3 ): d = 10.6-10.6 (CH 3 ), 20.7-20.8 (CH 3 -acetyl), 23.1 (CH 2 ), 28.0-28.4 ((CH 3 ) 3 -Boc), 29.7 (CH), 39.0-39.4 (CH), 42.4 (C6), 49.8 (CH 2- triazole), 50.4 (CH 2- N (Boc)), 69.7-70.8 (C5, C2, C3, C4), 79.7 (C (CH 3 ) 3 -Boc), 95.6 (C1), 124.7-125.3 (CH-triazole), 145.3 (= CN-triazole), 155.5-155.9 (C = O-Boc), 169.69-170.2 (C = O-acetyl)
FT-IR: (KBr, cm -1 ) 782.958, 878.417, 949.77, 1045.23, 1168.65, 1242.9, 1367.28, 1416.46, 1463.71, 1689.34, 1758.76, 2877.27, 2934.16, 2965.02, 3434.6
Calcd.for C 192 H 304 N 32 O 64 + 2.0 H 2 O: C 55.96, H 7.53, N 10.88. Found: C 55.95, H 7.58, N 10.64
(Deprotection of tertiary butoxycarbonyl group)
To 18.4 mg (4.50 × 10 −6 mol) of the above click reaction product, 500 ml of trifluoroacetic acid was added and dissolved. Then, it stirred for 20 minutes and obtained 18.9 mg (yield 100%) of white solid by depressurizingly distilling.
TLC Rf＝0.05 (ヘキサン/酢酸エチル=1/9)
1H NMR (400 MHz, DMSO-d6): δ= 0.82-0.84 (CH3), 1.22-1.27 (CH2), 1.52-1.56 (CH), 1.98-2.07 (CO-acetyl), 2.77 (CH2), 3.50-3.70 (H-4), 4.19-4.32 (CH2) 4.44 (H-5), 4.61-4.64 (H-6), 4.76 (H-3), 5.27-5.31 (H-3), 5.39 (H-1), 8.19 ( H-triazole), 9.20 (NH)
(アセチル基の脱保護)
上記の第三ブトキシカルボニル基を脱保護された化合物18.9 mg(4.50×10-3 mol)にメタノール1 mlを加えて溶解させた。続いて、ナトリウムメトキシドを1.94 mg (3.6×10-2 mol)加え、室温で4時間攪拌した。その後、陽イオン交換樹脂をpH 7になるまで加えた。陽イオン交換樹脂を濾去し、減圧留去により実施例２のシクロデキストリン誘導体（ｂ）を白色固体12.1 mg(収率76.1 %)として得た。
TLC R f = 0.05 (hexane / ethyl acetate = 1/9)
1 H NMR (400 MHz, DMSO-d 6 ): δ = 0.82-0.84 (CH 3 ), 1.22-1.27 (CH 2 ), 1.52-1.56 (CH), 1.98-2.07 (CO-acetyl), 2.77 (CH 2 ), 3.50-3.70 (H-4), 4.19-4.32 (CH 2 ) 4.44 (H-5), 4.61-4.64 (H-6), 4.76 (H-3), 5.27-5.31 (H-3) , 5.39 (H-1), 8.19 (H-triazole), 9.20 (NH)
(Deprotection of acetyl group)
To 18.9 mg (4.50 × 10 −3 mol) of a compound in which the third butoxycarbonyl group was deprotected, 1 ml of methanol was added and dissolved. Subsequently, 1.94 mg (3.6 × 10 −2 mol) of sodium methoxide was added, and the mixture was stirred at room temperature for 4 hours. Thereafter, cation exchange resin was added until pH 7 was reached. The cation exchange resin was removed by filtration, and the cyclodextrin derivative (b) of Example 2 was obtained as a white solid 12.1 mg (yield 76.1%) by distillation under reduced pressure.
TLC Rf＝0.57 (アンモニア/水/1-プロパノール/酢酸エチル=3/2/7/7)
1H NMR (400 MHz, DMSO-d6): δ=0.76-0.79 (CH3), 1.23-1.32 (CH2), 1.55-1.58 (CH), 2.81 (CH2), 3.50-3.70 (H-4, H-5, H-6), 3.68-3.70 (H-2), 4.12 (CH2), 4.47 (H-3), 5.11 (H-1),6.07-6.10 (OH), 8.09 (triazole) 9.12 (NH)
MS: [M+Na]+Calcd. for C120H208N32NaO32: 2632.5530, Found: 2632.5270, [M+K]+ Calcd. for C120H208KN32O32: 2648.5269, Found: 2648.5031
（実施例３）
シクロデキストリン誘導体（ｃ）の合成
(化合物[４]を用いたクリック反応)
硫酸銅 10.2 mg (4.06×10-5mol)を250 μlの純水に溶かし、アスコルビン酸ナトリウム100.6 mg(5.07×10-4 mol)を250 μlの純水に溶かしたものを加えた。その溶液を、DMSO 2 mlに溶かした化合物[４] 122 mg(5.62×10-5 mol)に加えた。ここにDMSO 2 mlに溶かした第三ブトキシカルボニル化２−メチルプロピルアミノプロピン107.1 mg(5.07×10-4 mol)を加えた。これにマイクロ波を照射し加熱(90 min, 120℃)した後、酢酸エチルに溶かし、これを5 %EDTA二ナトリウム水溶液で洗浄し、減圧留去にて溶媒を除去した残渣をシリカゲルカラム (塩化メチレン/メタノール) により精製し、白色固体107.6 mg(収率49.6 %)を得た。
TLC Rf＝0.48 (塩化メチレン/メタノール=20/1)
1H NMR (400 MHz, CDCl3): δ= 0.84 (CH3), 1.41 (C(CH3)3), 1.95 (CH), 2.04-2.09 (COCH3), 2.96-3.06 (CH2), 3.48-3.57(H-4), 4.39-4.55 (H-6,H-5), 4.66-4.68 (H-2), 4.86-4.89 (CH2), 5.38 (H-3), 5.62 (H-1), 7.68-7.73(triazole)
13C NMR (100 MHz, CDCl3): d = 20.0 (CH3), 20.7-21.9 (CH3-acetyl), 27.2-27.5 (CH2), 28.4 ((CH3)3-Boc), 29.70 (CH), 42.5-42.9 (CH), 49.4 (C6), 54.4 (CH2-triazole), 54.75 (CH2-N(Boc)), 70.9-70.6 (C5, C2, C3, C4), 79.7 (OC(CH3)3-Boc)], 95.7 (C1), 125.4 (CH2-triazole), 145.3 (=C-N-triazole), 155.9 (C=O-Boc), 169.6-170.2 (C=O-acetyl)
FT-IR :(KBr, cm-1) 408.835, 457.047, 599.753, 784.886, 1045.23, 1167.69, 1246.75, 1367.28, 1416.46, 1465.63, 1688.37, 1756.83, 2964.05, 3460.63
MS: [M+Na]+Calcd. for C176H272NaN32O64: 3880.9, Found: 3880.4
(第三ブトキシカルボニル基の脱保護)
上記のクリック反応生成物61.0 mg(1.58×10-5 mol)にトリフルオロ酢酸6 mlを加えて溶解させた。その後、30分攪拌し、減圧留去により白色固体62.7 mg(収率100.0 %)を得た。
TLC R f = 0.57 (ammonia / water / 1-propanol / ethyl acetate = 3/2/7/7)
1 H NMR (400 MHz, DMSO-d 6 ): δ = 0.76-0.79 (CH 3 ), 1.23-1.32 (CH 2 ), 1.55-1.58 (CH), 2.81 (CH 2 ), 3.50-3.70 (H- 4, H-5, H-6), 3.68-3.70 (H-2), 4.12 (CH 2 ), 4.47 (H-3), 5.11 (H-1), 6.07-6.10 (OH), 8.09 (triazole ) 9.12 (NH)
MS: [M + Na] + Calcd. For C 120 H 208 N 32 NaO 32 : 2632.5530, Found: 2632.5270, [M + K] + Calcd. For C 120 H 208 KN 32 O 32 : 2648.5269, Found: 2648.5031
(Example 3)
Synthesis of cyclodextrin derivative (c)
(Click reaction using compound [4])
Copper sulfate 10.2 mg (4.06 × 10 −5 mol) dissolved in 250 μl of pure water and sodium ascorbate 100.6 mg (5.07 × 10 −4 mol) dissolved in 250 μl of pure water were added. The solution was added to 122 mg (5.62 × 10 −5 mol) of the compound [4] dissolved in 2 ml of DMSO. To this was added 107.1 mg (5.07 × 10 −4 mol) of tert-butoxycarbonylated 2-methylpropylaminopropyne dissolved in 2 ml of DMSO. This was irradiated with microwaves and heated (90 min, 120 ° C), dissolved in ethyl acetate, washed with 5% aqueous EDTA disodium solution, and the solvent was removed by distillation under reduced pressure. Methylene / methanol) to obtain 107.6 mg (yield 49.6%) of a white solid.
TLC R f = 0.48 (methylene chloride / methanol = 20/1)
1 H NMR (400 MHz, CDCl 3 ): δ = 0.84 (CH 3 ), 1.41 (C (CH 3 ) 3 ), 1.95 (CH), 2.04-2.09 (COCH 3 ), 2.96-3.06 (CH 2 ), 3.48-3.57 (H-4), 4.39-4.55 (H-6, H-5), 4.66-4.68 (H-2), 4.86-4.89 (CH 2 ), 5.38 (H-3), 5.62 (H- 1), 7.68-7.73 (triazole)
13 C NMR (100 MHz, CDCl 3 ): d = 20.0 (CH 3 ), 20.7-21.9 (CH 3 -acetyl), 27.2-27.5 (CH 2 ), 28.4 ((CH 3 ) 3 -Boc), 29.70 ( CH), 42.5-42.9 (CH), 49.4 (C6), 54.4 (CH 2- triazole), 54.75 (CH 2- N (Boc)), 70.9-70.6 (C5, C2, C3, C4), 79.7 (OC (CH 3 ) 3 -Boc)], 95.7 (C1), 125.4 (CH 2- triazole), 145.3 (= CN-triazole), 155.9 (C = O-Boc), 169.6-170.2 (C = O-acetyl)
FT-IR: (KBr, cm -1 ) 408.835, 457.047, 599.753, 784.886, 1045.23, 1167.69, 1246.75, 1367.28, 1416.46, 1465.63, 1688.37, 1756.83, 2964.05, 3460.63
MS: [M + Na] + Calcd. For C 176 H 272 NaN 32 O 64 : 3880.9, Found: 3880.4
(Deprotection of tertiary butoxycarbonyl group)
To 61.0 mg (1.58 × 10 −5 mol) of the above click reaction product, 6 ml of trifluoroacetic acid was added and dissolved. Then, it stirred for 30 minutes and obtained 62.7 mg (yield 100.0%) of white solid by depressurizingly distilling.
TLC Rf＝0.15 (ヘキサン/酢酸エチル=1/9)
1H NMR (400 MHz, DMSO-d6): δ=0.85-0.88 (CH3), 1.88-1.96(CO-acetyl, CH), 2.50-2.73 (CH2), 3.57 (H-4), 4.18 (H-6), 4.35 (H-5), 4.64-4.71 (H-2, CH2), 5.28-5.03 (H-3), 5.38 (H-1), 8.14-8.25 ( H-triazole), 9.21 (NH)
(アセチル基の脱保護)
上記の第三ブトキシカルボニル基を脱保護された化合物30.0 mg(7.58×10-6 mol)にメタノール2 mlを加えて溶解させた。続いて、ナトリウムメトキシド3.28 mg(6.06×10-5 mol)を加え、室温で8時間攪拌した。その後、陽イオン交換樹脂をpH 7になるまで加えた。陽イオン交換樹脂を濾去し、減圧留去により実施例３の化合物（ｃ）を白色固体22.9 mg(収率91.6 %)として得た。
TLC Rf＝0.15 (アンモニア/水/1-プロパノール/酢酸エチル=0.5/2/5/3)、
1H NMR (400 MHz, DMSO-d6,): δ= 0.88-0.89 (CH3), 1.90-1.93 (CH), 2.72 (CH2), 3.22-3.50 (H-4, H-5, H-6), 3.68-3.70 (H-2), 4.11 (CH2), 4.43 (H-3), 5.11 (H-1), 6.08-6.12 (OH]) 8.08 (triazole), 9.15 (NH).
（実施例４）
シクロデキストリン誘導体（ｄ）の合成
(クリック反応)
硫酸銅 45.6 mg(2.30×10-4 mol)を100 μlの純水に溶かし、アスコルビン酸ナトリウム4.58 mg(1.84×10-4 mol)を100 μlの純水に溶かしたものを加えた。その溶液をDMSO 1 mlに溶かした化合物[４] 50 mg(2.30×10-5 mol)に加えた。ここにDMSO 1 mlに溶かした第三ブトキシカルボニル化シクロブチルメチルアミノプロピン 51.3 mg(2.30×10-4 mol)を加えた。これにマイクロ波を照射し加熱(30 min, 120℃)した。さらに硫酸銅45.6 mg(2.30×10-4 mol)を100 μlの純水に溶かし、アスコルビン酸ナトリウム4.58 mg(1.84×10-4 mol)を100 μlの純水に溶かしたものを加え、続いてDMSO 0.5 mlに溶かした第三ブトキシカルボニル化シクロブチルメチルアミノプロピン20.3 mg(9.10×10-5mol)を加えた。マイクロ波を照射して加熱(10 min, 120℃) した後、酢酸エチルに溶かし、これを5 %EDTA二ナトリウム水溶液で洗浄し、減圧留去にて溶媒を除去した残渣をシリカゲルカラム(塩化メチレン/メタノール)により精製し、白色固体71.5 mg (収率78.7 %)を得た。
TLC Rf＝0.20 (塩化メチレン/メタノール=20/1 (v/v))
1H NMR (400 MHz, CDCl3): δ= 1.42 [C(CH3)3], 1.67 [CH2], 1.80-1.94 [CH2], 1.93 [CH2], 2.00-2.05 [CH3], 2.55 [CH2], 3.23-3.34 [CH2], 3.55-3.61 [CH], 4.34-4.48 [CH, CH2], 4.67-4.69 [CH], 4.86 [CH2], 5.36-5.40 [CH], 5.64 [CH], 7.69-7.71 [NH]
FT-IR (KBr, cm-1) 462.83, 601.68, 785.85, 881.31, 1046.19, 1145.51, 1168.65, 1245.79, 1368.25, 1413.57, 1458.89, 1625,70, 1692.23, 1757.80, 2974.66, 3446.17
(アセチル基の脱保護)
上記のクリック反応生成物40.0 mg(1.01×10-5 mol)にメタノール3 mlを加えて溶解させた。続いて、ナトリウムメトキシドをpH 9になるまで加え、室温で12時間攪拌した。その後、陽イオン交換樹脂をpH 7になるまで加えた。陽イオン交換樹脂を濾去し、減圧留去により白色固体35.1 mg (収率100 %)を得た。
TLC Rf＝0.80 (アンモニア/水/1-プロパノール/酢酸エチル=3/2/7/7)
1H NMR (400 MHz, DMSO-d6): δ= 1.23 [C(CH3)3], 1.34 [CH2], 1.47 [CH2], 1.57 [CH2], 3.17-3.24 [CH2, CH2], 3.64-3.66 [CH], 4.03 [CH], 4.14 [CH], 4.35 [CH2], 5.11 [CH], 5.94 [OH], 7.69-7.71 [NH]
(第三ブトキシカルボニル基の脱保護)
アセチル基を脱保護された化合物30.3 mg(9.23×10-6 mol)にトリフルオロ酢酸1 mlを加えて溶解させた。その後、30分攪拌し、減圧留去により実施例４の化合物（ｄ）を白色固体33.4 mg (収率100 %)として得た。
TLC Rf＝0.35 (アンモニア/水/1-プロパノール/酢酸エチル=3/2/7/7 )
1H NMR (400 MHz, DMSO-d6): δ= 1.73 [CH2], 2.50-2.57 [CH], 2.95 [CH2], 3.39-3.40 [CH2], 3.68-3.70 [CH], 4.10 [CH], 4.45-4.53 [CH2], 5.11 [CH], 6.08 [OH], 8.08 [NH], 9.10 [NH2]
（実施例５）
シクロデキストリン誘導体（ｅ）の合成
(クリック反応)
硫酸銅 4.57 mg(1.85×10-5 mol)を160 μlの純水に溶かし、アスコルビン酸ナトリウム45.2 mg(2.31×10-4 mol)を140 μlの純水に溶かしたものを加えた。その溶液をDMSO 1 mlに溶かした化合物[４] 50 mg(2.30×10-5 mol)に加えた。ここにDMSO1.3 mlに溶かした第三ブトキシカルボニル化シクロペンチルメチルアミノプロピン 54.6 mg(2.30×10-4 mol)を加えた。これにマイクロ波を照射し加熱(10 min, 120℃) した後、酢酸エチルに溶かし、これを5 %EDTA二ナトリウム水溶液で洗浄し、減圧留去にて溶媒を除去した残渣をシリカゲルカラム（ヘキサン/酢酸エチル） により精製し、白色固体60.1 mg(収率64.2 %)を得た。
TLC Rf＝0.66 (ヘキサン/酢酸エチル=1/9 (v/v))
1H NMR (400 MHz, CDCl3): δ= 1.19 [CH2], 1.41 [C(CH3)3], 1.51 [CH2], 1.61 [CH2], 2.04-2.05 [COCH3], 2.17-2.21 [CH], 3.14-3.20 [CH2], 3.57 [H-4], 4.38-4.48 [H-5, H-6], 4.67-4.69 [H-2], 4.86 [CH2], 5.38 [H-3], 5.64 [H-1], 7.67-7.72 [triazole]
13C NMR (100 MHz, CDCl3): d = 20.7-20.9 [CH3CO], 24.9 [CH2], 28.4 [(CH3)3-Boc], 29.7-30.1 [CH2], 38.9 [CH], 42.1-42.5 [C6], 49.7 (CH2-triazole), 51.6 (CH2-N(Boc)), 69.6-70.7 [C5, C2, C3, C4], 79.6 [OC(CH3)3-Boc], 95.6 [C1], 124.7-125.3 [CH-triazol], 145.5 [=CN-triazol], 155.5-155.7 [C=O-Boc ], 169.6-170.2 [C=O-acetyl]
FT-IR :(KBr, cm-1) 460.904, 524.543, 566.005, 601.682, 647.965, 733.782, 782.958, 822.491, 879.381, 949.77, 1045.23, 1161.9, 1245.79, 1367.28, 1415.49, 1461.78, 1545.67, 1691.27, 1758.76, 2869.56, 2953.45, 3148.22, 3480.88
Anal.: Calcd. for C192H288N32O64+ 2.0 H2O: C 56.18, H 7.17, N 10.92. Found: C 56.27, H 7.25, N 10.63
(第三ブトキシカルボニル基の脱保護)
上記のクリック反応生成物のうち、42.3 mg(1.04×10-2 mol)にトリフルオロ酢酸 1 mlを加えて溶解させた。その後、20分攪拌し、減圧留去により白色固体41.0 mg(収率94.3 %)を得た。
TLC R f = 0.15 (hexane / ethyl acetate = 1/9)
1 H NMR (400 MHz, DMSO-d 6 ): δ = 0.85-0.88 (CH 3 ), 1.88-1.96 (CO-acetyl, CH), 2.50-2.73 (CH 2 ), 3.57 (H-4), 4.18 (H-6), 4.35 (H-5), 4.64-4.71 (H-2, CH 2 ), 5.28-5.03 (H-3), 5.38 (H-1), 8.14-8.25 (H-triazole), 9.21 (NH)
(Deprotection of acetyl group)
To 30.0 mg (7.58 × 10 −6 mol) of a compound in which the third butoxycarbonyl group was deprotected, 2 ml of methanol was added and dissolved. Subsequently, 3.28 mg (6.06 × 10 −5 mol) of sodium methoxide was added, and the mixture was stirred at room temperature for 8 hours. Thereafter, cation exchange resin was added until pH 7 was reached. The cation exchange resin was removed by filtration, and the compound (c) of Example 3 was obtained as a white solid 22.9 mg (yield 91.6%) by distillation under reduced pressure.
TLC R f = 0.15 (ammonia / water / 1-propanol / ethyl acetate = 0.5 / 2/5/3),
1 H NMR (400 MHz, DMSO-d 6, ): δ = 0.88-0.89 (CH 3 ), 1.90-1.93 (CH), 2.72 (CH 2 ), 3.22-3.50 (H-4, H-5, H -6), 3.68-3.70 (H-2), 4.11 (CH 2 ), 4.43 (H-3), 5.11 (H-1), 6.08-6.12 (OH)) 8.08 (triazole), 9.15 (NH).
Example 4
Synthesis of cyclodextrin derivative (d)
(Click reaction)
Copper sulfate 45.6 mg (2.30 × 10 −4 mol) was dissolved in 100 μl of pure water, and sodium ascorbate 4.58 mg (1.84 × 10 −4 mol) dissolved in 100 μl of pure water was added. The solution was added to 50 mg (2.30 × 10 −5 mol) of the compound [4] dissolved in 1 ml of DMSO. To this was added 51.3 mg (2.30 × 10 −4 mol) of tert-butoxycarbonylated cyclobutylmethylaminopropyne dissolved in 1 ml of DMSO. This was irradiated with microwaves and heated (30 min, 120 ° C.). Add 45.6 mg (2.30 × 10 -4 mol) of copper sulfate in 100 μl of pure water, add 4.58 mg (1.84 × 10 -4 mol) of sodium ascorbate in 100 μl of pure water, and then add 20.3 mg (9.10 × 10 −5 mol) of tert-butoxycarbonylated cyclobutylmethylaminopropyne dissolved in 0.5 ml of DMSO was added. After heating by microwave irradiation (10 min, 120 ° C), the residue was dissolved in ethyl acetate, washed with 5% disodium EDTA aqueous solution, and the solvent was removed by distillation under reduced pressure. / Methanol) to obtain 71.5 mg (yield 78.7%) of a white solid.
TLC R f = 0.20 (methylene chloride / methanol = 20/1 (v / v))
1 H NMR (400 MHz, CDCl 3 ): δ = 1.42 [C (CH 3 ) 3 ], 1.67 [CH 2 ], 1.80-1.94 [CH 2 ], 1.93 [CH 2 ], 2.00-2.05 [CH 3 ] , 2.55 [CH 2 ], 3.23-3.34 [CH 2 ], 3.55-3.61 [CH], 4.34-4.48 [CH, CH 2 ], 4.67-4.69 [CH], 4.86 [CH 2 ], 5.36-5.40 [CH ], 5.64 [CH], 7.69-7.71 [NH]
FT-IR (KBr, cm -1 ) 462.83, 601.68, 785.85, 881.31, 1046.19, 1145.51, 1168.65, 1245.79, 1368.25, 1413.57, 1458.89, 1625,70, 1692.23, 1757.80, 2974.66, 3446.17
(Deprotection of acetyl group)
To 40.0 mg (1.01 × 10 −5 mol) of the above click reaction product, 3 ml of methanol was added and dissolved. Subsequently, sodium methoxide was added until pH 9 and stirred at room temperature for 12 hours. Thereafter, cation exchange resin was added until pH 7 was reached. The cation exchange resin was removed by filtration, and 35.1 mg (yield 100%) of a white solid was obtained by evaporation under reduced pressure.
TLC R f = 0.80 (ammonia / water / 1-propanol / ethyl acetate = 3/2/7/7)
1 H NMR (400 MHz, DMSO-d 6 ): δ = 1.23 [C (CH 3 ) 3 ], 1.34 [CH 2 ], 1.47 [CH 2 ], 1.57 [CH 2 ], 3.17-3.24 [CH 2 , CH 2 ], 3.64-3.66 [CH], 4.03 [CH], 4.14 [CH], 4.35 [CH 2 ], 5.11 [CH], 5.94 [OH], 7.69-7.71 [NH]
(Deprotection of tertiary butoxycarbonyl group)
To 30.3 mg (9.23 × 10 −6 mol) of the compound in which the acetyl group was deprotected, 1 ml of trifluoroacetic acid was added and dissolved. Thereafter, the mixture was stirred for 30 minutes, and the compound (d) of Example 4 was obtained as 33.4 mg (yield 100%) of a white solid by distillation under reduced pressure.
TLC R f = 0.35 (ammonia / water / 1-propanol / ethyl acetate = 3/2/7/7)
1 H NMR (400 MHz, DMSO-d 6 ): δ = 1.73 [CH 2 ], 2.50-2.57 [CH], 2.95 [CH 2 ], 3.39-3.40 [CH 2 ], 3.68-3.70 [CH], 4.10 [CH], 4.45-4.53 [CH 2 ], 5.11 [CH], 6.08 [OH], 8.08 [NH], 9.10 [NH 2 ]
(Example 5)
Synthesis of cyclodextrin derivative (e)
(Click reaction)
Copper sulfate 4.57 mg (1.85 × 10 −5 mol) was dissolved in 160 μl of pure water, and sodium ascorbate 45.2 mg (2.31 × 10 −4 mol) dissolved in 140 μl of pure water was added. The solution was added to 50 mg (2.30 × 10 −5 mol) of the compound [4] dissolved in 1 ml of DMSO. To this was added 54.6 mg (2.30 × 10 −4 mol) of tert-butoxycarbonylated cyclopentylmethylaminopropyne dissolved in 1.3 ml of DMSO. This was irradiated with microwaves and heated (10 min, 120 ° C), dissolved in ethyl acetate, washed with 5% aqueous disodium EDTA solution, and the solvent was removed by distillation under reduced pressure. / Ethyl acetate) to obtain 60.1 mg (yield 64.2%) of a white solid.
TLC R f = 0.66 (hexane / ethyl acetate = 1/9 (v / v))
1 H NMR (400 MHz, CDCl 3 ): δ = 1.19 [CH 2 ], 1.41 [C (CH 3 ) 3 ], 1.51 [CH 2 ], 1.61 [CH 2 ], 2.04-2.05 [COCH 3 ], 2.17 -2.21 [CH], 3.14-3.20 [CH 2 ], 3.57 [H-4], 4.38-4.48 [H-5, H-6], 4.67-4.69 [H-2], 4.86 [CH 2 ], 5.38 [H-3], 5.64 [H-1], 7.67-7.72 [triazole]
13 C NMR (100 MHz, CDCl 3 ): d = 20.7-20.9 [CH 3 CO], 24.9 [CH 2 ], 28.4 [(CH 3 ) 3 -Boc], 29.7-30.1 [CH 2 ], 38.9 [CH ], 42.1-42.5 [C6], 49.7 (CH 2- triazole), 51.6 (CH 2- N (Boc)), 69.6-70.7 [C5, C2, C3, C4], 79.6 [OC (CH 3 ) 3- Boc], 95.6 [C1], 124.7-125.3 [CH-triazol], 145.5 [= CN-triazol], 155.5-155.7 [C = O-Boc], 169.6-170.2 [C = O-acetyl]
FT-IR: (KBr, cm -1 ) 460.904, 524.543, 566.005, 601.682, 647.965, 733.782, 782.958, 822.491, 879.381, 949.77, 1045.23, 1161.9, 1245.79, 1367.28, 1415.49, 1461.78, 1545.67, 1691.27, , 2953.45, 3148.22, 3480.88
Anal .: Calcd. For C 192 H 288 N 32 O 64 + 2.0 H 2 O: C 56.18, H 7.17, N 10.92. Found: C 56.27, H 7.25, N 10.63
(Deprotection of tertiary butoxycarbonyl group)
Of the above click reaction product, 1 ml of trifluoroacetic acid was dissolved in 42.3 mg (1.04 × 10 −2 mol). Thereafter, the mixture was stirred for 20 minutes, and 41.0 mg (yield 94.3%) of a white solid was obtained by distillation under reduced pressure.
TLC Rf＝0.05 (ヘキサン/酢酸エチル=1/9 (v/v)、ナフトレゾルシン発色)
1H NMR (400 MHz, DMSO-d6): δ=1.121-1.134 [CH2], 1.459-1.531 [CH2], 1.699-1.714 [CH2], 1.995 [COCH3], 2.065-2.103 [CH], 2.833 [CH2], 3.446-3.569 [H-4, CH2], 4.181 [H-6], 4.427 [H-5], 4.634-4.656 [H-2], 4.735 [CH2], 5.276-5.321[H-3], 5.380 [H-1], 8.186[H-triazol], 9.244[NH]
(アセチル基の脱保護)
上記の第三ブトキシカルボニル基を脱保護された化合物 28.2 mg(6.75×10-3mol)にメタノール1 mlを加えて溶解させた。続いて、ナトリウムメトキシドを2.91 mg(5.4×10-2 mol)加え、室温で4時間攪拌した。その後、陽イオン交換樹脂をpH 7になるまで加えた。陽イオン交換樹脂を濾去し、減圧留去により実施例５の化合物（ｅ）を白色固体18.1 mg(収率76.7 %)として得た。
TLC Rf＝0.15 (アンモニア/水/1-プロパノール/酢酸エチル=3/2/7/7)
1H NMR (400 MHz, DMSO-d6,): δ= 1.149-1.233 [CH2], 1.475-1.547 [CH2], 1.722 [CH2], 2.083-2.103 [CH], 2.859 [CH2], 3.219-3.394 [H-4, H-6], 3.698 [H-5], 4.098 [CH2, H-2], 4.433 [H-3], 5.106 [H-1], 6.099 [OH], 8.080[triazole], 9.193 [NH]
MS: [M+H]+ Calcd. for C120H193N32O32: 2594.4546, Found: 2594.4702
[M+Na]+Calcd. for C120H192N32NaO32: 2616.4278, Found: 2616.4516
（評価）
上記のようにして合成した実施例１〜５のシクロデキストリン誘導体（ａ）〜（ｅ）の抗菌性を最小発育濃度（ＭＩＣ）で評価した。評価にはグラム陽性菌として黄色ブドウ球菌および枯草菌、グラム陰性菌として大腸菌、ネズミチフス菌、緑膿菌を使用した。
TLC R f = 0.05 (Hexane / Ethyl acetate = 1/9 (v / v), Naphtresolcin coloring)
1 H NMR (400 MHz, DMSO-d 6 ): δ = 1.121-1.134 [CH 2 ], 1.459-1.531 [CH 2 ], 1.699-1.714 [CH 2 ], 1.995 [COCH 3 ], 2.065-2.103 [CH ], 2.833 [CH 2 ], 3.446-3.569 [H-4, CH 2 ], 4.181 [H-6], 4.427 [H-5], 4.634-4.656 [H- 2 ], 4.735 [CH 2 ], 5.276 -5.321 [H-3], 5.380 [H-1], 8.186 [H-triazol], 9.244 [NH]
(Deprotection of acetyl group)
To 28.2 mg (6.75 × 10 −3 mol) of a compound in which the third butoxycarbonyl group was deprotected, 1 ml of methanol was added and dissolved. Subsequently, 2.91 mg (5.4 × 10 −2 mol) of sodium methoxide was added, and the mixture was stirred at room temperature for 4 hours. Thereafter, cation exchange resin was added until pH 7 was reached. The cation exchange resin was removed by filtration, and the compound (e) of Example 5 was obtained as a white solid (18.1 mg, yield 76.7%) by distillation under reduced pressure.
TLC R f = 0.15 (ammonia / water / 1-propanol / ethyl acetate = 3/2/7/7)
1 H NMR (400 MHz, DMSO-d 6, ): δ = 1.149-1.233 [CH 2 ], 1.475-1.547 [CH 2 ], 1.722 [CH 2 ], 2.083-2.103 [CH], 2.859 [CH 2 ] , 3.219-3.394 [H-4, H-6], 3.698 [H-5], 4.098 [CH 2, H-2], 4.433 [H-3], 5.106 [H-1], 6.099 [OH], 8.080 [triazole], 9.193 [NH]
MS: [M + H] + Calcd. For C 120 H 193 N 32 O 32 : 2594.4546, Found: 2594.4702
[M + Na] + Calcd. For C 120 H 192 N 32 NaO 32 : 2616.4278, Found: 2616.4516
(Evaluation)
The antibacterial properties of the cyclodextrin derivatives (a) to (e) of Examples 1 to 5 synthesized as described above were evaluated at the minimum growth concentration (MIC). For the evaluation, Staphylococcus aureus and Bacillus subtilis were used as Gram-positive bacteria, and Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa were used as Gram-negative bacteria.
評価方法は以下の通りである。 The evaluation method is as follows.
すなわち、細菌は、一般細菌用乾燥ブイヨンあるいは大腸菌はポリペプトンを加えた最小塩培地で培養し、対数増殖期の状態のものを使用した。最小発育濃度は、Ｍｕｌｌｅｒ Ｈｉｎｔｏｎ培地、菌液、各濃度の候補化合物溶液を、３７℃で２０時間、培養することにより、試験化合物がその細菌の生育を阻止した最小濃度とした。なお初期の菌濃度は１ｘ１０４ 細胞/ｍＬとした。 That is, bacteria were cultured in a dry bouillon for general bacteria or Escherichia coli was cultured in a minimal salt medium to which polypeptone was added, and those in a logarithmic growth phase were used. The minimum growth concentration was determined as the minimum concentration at which the test compound inhibited the growth of the bacteria by culturing the Muller Hinton medium, the bacterial solution, and each candidate compound solution at 37 ° C. for 20 hours. The initial bacterial concentration was 1 × 10 4 cells / mL.
最小発育濃度（ＭＩＣ）で評価の結果を表１に示す。 Table 1 shows the results of the evaluation based on the minimum growth concentration (MIC).
一般に最小発育濃度が１０００μｇ/mLを上回る場合には、その物質には抗菌性はないと判断され、１０〜１０００μｇ/ｍLの場合には中程度、１０μｇ/ｍL以下では強力な抗菌性ありとされる。 In general, when the minimum growth concentration exceeds 1000 μg / mL, it is judged that the substance is not antibacterial. When the concentration is 10 to 1000 μg / mL, it is moderate, and at 10 μg / mL or less, it is considered to have strong antibacterial properties. The
上記表１に示すように、末端が鎖上のアルキル部である２−メチルプロピルアミノメチルトリアゾール基を持つ実施例３の化合物（Ｃ）は、黄色ブドウ球菌に対して６４μｇ/ｍLという中程度の抗菌性を示した。同じく末端が鎖上のアルキル部である２−エチルブチルアミノメチルトリアゾール基を持つ実施例２の化合物（ｂ）は、グラム陽性菌である黄色ブドウ球菌および枯草菌、グラム陰性菌である大腸菌およびネズミチフス菌に対しては４または８μｇ/ｍLという強力な抗菌性を示し、緑膿菌に対しても６４μｇ/ｍLという中程度の抗菌性を示した。 As shown in Table 1 above, the compound (C) of Example 3 having a 2-methylpropylaminomethyltriazole group whose terminal is an alkyl moiety on the chain is a medium of 64 μg / mL against Staphylococcus aureus. It showed antibacterial properties. Similarly, the compound (b) of Example 2 having a 2-ethylbutylaminomethyltriazole group whose terminal is an alkyl part on the chain is a gram-positive bacterium, Staphylococcus aureus and Bacillus subtilis, a gram-negative bacterium, Escherichia coli and Salmonella typhimurium. It showed strong antibacterial activity of 4 or 8 μg / mL against bacteria and moderate antibacterial activity against Pseudomonas aeruginosa of 64 μg / mL.
一方、末端が環状のアルキル部である実施例１、４、５の化合物（ａ）、（ｄ）、（ｅ）については、グラム陽性菌である黄色ブドウ球菌および枯草菌、グラム陰性菌である大腸菌およびネズミチフス菌に対しては、いずれも抗菌性を示した。すなわち、シクロへキシル部を持つ実施例１の化合物（ａ）はグラム陰性菌である大腸菌およびネズミチフス菌に対する最小発育濃度はいずれも１６μｇ/ｍLであり、グラム陽性菌には８μｇ/ｍLという強力な抗菌性を示した。また、シクロブチル部を持つ実施例４の化合物（ｄ）は、ネズミチフス菌に対する最小発育濃度は３２μｇ/ｍLであるものの、グラム陰性菌である大腸菌、およびグラム陽性菌には８μｇ/ｍLという強力な抗菌性を示した。さらに、シクロペンチル部を持つ実施例５の化合物（ｅ）は、黄色ブドウ球菌、枯草菌、大腸菌およびネズミチフス菌の全てに対して４または８μｇ/ｍLという強力な抗菌性を示した。なお、グラム陰性菌である緑膿菌への最小発育濃度はいずれも１２８μｇ/ｍL超であった。 On the other hand, the compounds (a), (d), and (e) of Examples 1, 4, and 5 having a cyclic alkyl moiety at the end are Gram-positive bacteria, Staphylococcus aureus, Bacillus subtilis, and Gram-negative bacteria. Both showed antibacterial activity against Escherichia coli and Salmonella typhimurium. That is, the compound (a) of Example 1 having a cyclohexyl moiety has a minimum growth concentration of 16 μg / mL for Escherichia coli and Salmonella typhimurium, which are Gram-negative bacteria, and is a strong 8 μg / mL for Gram-positive bacteria. It showed antibacterial properties. In addition, the compound (d) of Example 4 having a cyclobutyl moiety has a strong antibacterial activity of 8 μg / mL for Escherichia coli and Gram-positive bacteria, although the minimum growth concentration against Salmonella typhimurium is 32 μg / mL. Showed sex. Furthermore, the compound (e) of Example 5 having a cyclopentyl moiety showed a strong antibacterial activity of 4 or 8 μg / mL against all of Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Salmonella typhimurium. Note that the minimum growth concentration for Pseudomonas aeruginosa, which is a gram-negative bacterium, was more than 128 μg / mL.
以上の結果から、γシクロデキストリンに疎水性アルキル部を持つアミノトリアゾール基を導入した化合物は、抗菌性を発現する上で有効であることが分かった。また、アルキル部が鎖状または環状のいずれでも抗菌性を示すことが分かった。さらには、ここで示された抗菌性はその分子設計から膜傷害機構に起因するものと合理的に推定され、シクロデキストリンを利用して抗菌性を発現する物質を構築する前述の分子設計とその調製方法の有効性を実証している。 From the above results, it was found that a compound in which an aminotriazole group having a hydrophobic alkyl moiety was introduced into γ-cyclodextrin was effective in developing antibacterial properties. It was also found that the alkyl part exhibits antibacterial properties regardless of whether it is linear or cyclic. Furthermore, the antibacterial properties shown here are reasonably presumed to be due to the membrane damage mechanism from the molecular design, and the molecular design described above for constructing a substance that exhibits antibacterial properties using cyclodextrin and its molecular design. The effectiveness of the preparation method is demonstrated.
そしてここでは、 親水性であるシクロデキストリンのグルコース部分にトリアゾール部を介してアルキルアミノメチル部を連結することで細菌への抗菌性を発現させた。これはグルコースの性質を利用して親水性と疎水性のバランスを適切に調節することにより抗菌物質ができることを示している。それゆえに、シクロデキストリンと同様にグルコースからなるオリゴ糖〜多糖にアルキルアミノアルキルトリアゾール部を導入することで抗菌物質が合成できると考えられる。さらに、グルコースの異性体で同様の性質を持つ単糖からなるオリゴ糖〜多糖からも抗菌物質が合成できることを合理的に推論できる。 And here, antibacterial activity against bacteria was expressed by connecting an alkylaminomethyl part to the glucose part of cyclodextrin, which is hydrophilic, via a triazole part. This indicates that an antibacterial substance can be produced by appropriately adjusting the balance between hydrophilicity and hydrophobicity using the properties of glucose. Therefore, it is considered that an antibacterial substance can be synthesized by introducing an alkylaminoalkyltriazole moiety into an oligosaccharide to polysaccharide consisting of glucose as in cyclodextrin. Furthermore, it can be reasonably inferred that an antibacterial substance can be synthesized from oligosaccharides to polysaccharides composed of monosaccharides having the same properties as isomers of glucose.
この発明は上記発明の実施の態様及び実施例の説明に何ら限定されるものではない。特許請求の範囲を逸脱せず、当業者が容易に想到できる範囲で種々の変形態様もこの発明に含まれる。 The present invention is not limited to the description of the embodiments and examples of the invention described above. Various modifications are also included in the present invention as long as those skilled in the art can easily conceive without departing from the scope of the claims.
本発明の糖誘導体又はその塩は抗菌剤として利用可能である。また、本発明の試薬は、物質を抗菌化する方法に利用可能である。 The sugar derivative of the present invention or a salt thereof can be used as an antibacterial agent. Moreover, the reagent of this invention can be utilized for the method of making a substance antibacterial.
Claims (20)
As a reagent for synthesizing an antibacterial agent, an azide of a sugar in which a monosaccharide represented by the following general formula is linked in a chain or cyclic form with a glycosidic bond (where the azide group is substituted with a primary hydroxyl group of the monosaccharide) N represents an integer of 2 or more).
(一般式中のアジド基は単糖の一級水酸基と置換している。ｎは２以上の整数を示す。)
（一般式中のR１はアルキレン基、R２は分岐してもよくシクロ環を有してもよいアルキル基を示し、R3は水素又は脱保護可能な置換基を示す。） 9. The azide of a sugar represented by the following general formula, in which monosaccharides are linked in a chain or cyclic form with a glycosidic bond, and a terminal alkyne represented by the following general formula, are reacted. A process for producing the sugar derivative or salt thereof according to claim 1.
(The azide group in the general formula is substituted with a primary hydroxyl group of a monosaccharide. N represents an integer of 2 or more.)
(In the general formula, R 1 represents an alkylene group, R 2 represents an alkyl group which may be branched or have a cyclo ring, and R 3 represents hydrogen or a deprotectable substituent.)
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| CN105085693A (en) * | 2015-07-16 | 2015-11-25 | 中国科学院烟台海岸带研究所 | 1,2,3-triazole starch derivatives, and preparation method and application thereof |
| JP2016053152A (en) * | 2014-07-25 | 2016-04-14 | 国立大学法人 名古屋工業大学 | Sugar derivative or salt thereof, antibacterial agent or antibacterial activity enhancer each using the same, reagent for synthesizing the same and method for producing the same using the reagent |
| WO2018051903A1 (en) * | 2016-09-13 | 2018-03-22 | 国立大学法人名古屋工業大学 | Sugar derivative or salt thereof, and antibacterial agent or antibacterial activity enhancer using same |
| JP2019031593A (en) * | 2017-08-04 | 2019-02-28 | 国立大学法人 名古屋工業大学 | Saccharide derivative, antibacterial agent and cosmetic |
| EP3380554B1 (en) | 2015-11-25 | 2020-06-10 | Fresenius Kabi iPSUM S.r.l. | Crystalline forms of per-chloro-gamma-cyclodextrines |
| EP3380530B1 (en) | 2015-11-25 | 2020-10-07 | Fresenius Kabi iPSUM S.r.l. | An improved process for the preparation of sugammadex and its intermediates |
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| JP2008528761A (en) * | 2005-01-28 | 2008-07-31 | ピナクル ファーマシューティカルズ，インク． | Β-cyclodextrin derivatives as antibacterial agents |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2016053152A (en) * | 2014-07-25 | 2016-04-14 | 国立大学法人 名古屋工業大学 | Sugar derivative or salt thereof, antibacterial agent or antibacterial activity enhancer each using the same, reagent for synthesizing the same and method for producing the same using the reagent |
| CN105085693A (en) * | 2015-07-16 | 2015-11-25 | 中国科学院烟台海岸带研究所 | 1,2,3-triazole starch derivatives, and preparation method and application thereof |
| EP3380554B1 (en) | 2015-11-25 | 2020-06-10 | Fresenius Kabi iPSUM S.r.l. | Crystalline forms of per-chloro-gamma-cyclodextrines |
| EP3380530B1 (en) | 2015-11-25 | 2020-10-07 | Fresenius Kabi iPSUM S.r.l. | An improved process for the preparation of sugammadex and its intermediates |
| WO2018051903A1 (en) * | 2016-09-13 | 2018-03-22 | 国立大学法人名古屋工業大学 | Sugar derivative or salt thereof, and antibacterial agent or antibacterial activity enhancer using same |
| JPWO2018051903A1 (en) * | 2016-09-13 | 2019-06-24 | 国立大学法人 名古屋工業大学 | Sugar derivative or salt thereof, antibacterial agent or antibacterial activity enhancer using the same |
| JP2019031593A (en) * | 2017-08-04 | 2019-02-28 | 国立大学法人 名古屋工業大学 | Saccharide derivative, antibacterial agent and cosmetic |
| JP7044313B2 (en) | 2017-08-04 | 2022-03-30 | 国立大学法人 名古屋工業大学 | Sugar derivatives, antibacterial agents and cosmetics |
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