KR20240036894A - Preparing method of Ectoine - Google Patents
Preparing method of Ectoine Download PDFInfo
- Publication number
- KR20240036894A KR20240036894A KR1020220115496A KR20220115496A KR20240036894A KR 20240036894 A KR20240036894 A KR 20240036894A KR 1020220115496 A KR1020220115496 A KR 1020220115496A KR 20220115496 A KR20220115496 A KR 20220115496A KR 20240036894 A KR20240036894 A KR 20240036894A
- Authority
- KR
- South Korea
- Prior art keywords
- ectoine
- present
- acid
- formula
- producing
- Prior art date
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- WQXNXVUDBPYKBA-YFKPBYRVSA-N ectoine Chemical compound CC1=[NH+][C@H](C([O-])=O)CCN1 WQXNXVUDBPYKBA-YFKPBYRVSA-N 0.000 title claims abstract description 70
- WQXNXVUDBPYKBA-UHFFFAOYSA-N Ectoine Natural products CC1=NCCC(C(O)=O)N1 WQXNXVUDBPYKBA-UHFFFAOYSA-N 0.000 title claims abstract description 61
- 238000000034 method Methods 0.000 title claims description 30
- 238000004519 manufacturing process Methods 0.000 claims abstract description 54
- -1 2-nosyl Chemical group 0.000 claims abstract description 30
- 150000001875 compounds Chemical class 0.000 claims abstract description 28
- OGNSCSPNOLGXSM-VKHMYHEASA-N L-2,4-diaminobutyric acid Chemical compound NCC[C@H](N)C(O)=O OGNSCSPNOLGXSM-VKHMYHEASA-N 0.000 claims abstract description 17
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 claims abstract description 10
- 229930182816 L-glutamine Natural products 0.000 claims abstract description 9
- 239000000203 mixture Substances 0.000 claims description 25
- 230000008569 process Effects 0.000 claims description 19
- 239000003153 chemical reaction reagent Substances 0.000 claims description 13
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 claims description 12
- 229910052740 iodine Inorganic materials 0.000 claims description 12
- 239000011630 iodine Substances 0.000 claims description 12
- 150000001356 alkyl thiols Chemical class 0.000 claims description 8
- 150000003573 thiols Chemical class 0.000 claims description 8
- 239000003960 organic solvent Substances 0.000 claims description 7
- 125000000217 alkyl group Chemical group 0.000 claims description 6
- CWERGRDVMFNCDR-UHFFFAOYSA-N thioglycolic acid Chemical compound OC(=O)CS CWERGRDVMFNCDR-UHFFFAOYSA-N 0.000 claims description 6
- OGNSCSPNOLGXSM-UHFFFAOYSA-N 2,4-diaminobutyric acid Chemical compound NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 claims description 4
- 125000004432 carbon atom Chemical group C* 0.000 claims description 2
- 239000002904 solvent Substances 0.000 abstract description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 6
- 125000001736 nosyl group Chemical group S(=O)(=O)(C1=CC=C([N+](=O)[O-])C=C1)* 0.000 abstract description 5
- 238000007363 ring formation reaction Methods 0.000 abstract description 5
- 238000006462 rearrangement reaction Methods 0.000 abstract description 3
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 abstract 2
- 239000000543 intermediate Substances 0.000 description 28
- 238000006243 chemical reaction Methods 0.000 description 25
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 24
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 18
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 18
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 13
- 239000000243 solution Substances 0.000 description 12
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 9
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 9
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 9
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 8
- 239000002253 acid Substances 0.000 description 8
- 239000007787 solid Substances 0.000 description 8
- 238000003756 stirring Methods 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- 150000003839 salts Chemical class 0.000 description 7
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 6
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 6
- ZBIKORITPGTTGI-UHFFFAOYSA-N [acetyloxy(phenyl)-$l^{3}-iodanyl] acetate Chemical compound CC(=O)OI(OC(C)=O)C1=CC=CC=C1 ZBIKORITPGTTGI-UHFFFAOYSA-N 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- RMVRSNDYEFQCLF-UHFFFAOYSA-N thiophenol Chemical compound SC1=CC=CC=C1 RMVRSNDYEFQCLF-UHFFFAOYSA-N 0.000 description 6
- FANCTJAFZSYTIS-IQUVVAJASA-N (1r,3s,5z)-5-[(2e)-2-[(1r,3as,7ar)-7a-methyl-1-[(2r)-4-(phenylsulfonimidoyl)butan-2-yl]-2,3,3a,5,6,7-hexahydro-1h-inden-4-ylidene]ethylidene]-4-methylidenecyclohexane-1,3-diol Chemical compound C([C@@H](C)[C@@H]1[C@]2(CCCC(/[C@@H]2CC1)=C\C=C\1C([C@@H](O)C[C@H](O)C/1)=C)C)CS(=N)(=O)C1=CC=CC=C1 FANCTJAFZSYTIS-IQUVVAJASA-N 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 5
- 238000002425 crystallisation Methods 0.000 description 5
- 230000008025 crystallization Effects 0.000 description 5
- 239000012153 distilled water Substances 0.000 description 5
- 150000007529 inorganic bases Chemical class 0.000 description 5
- 229910052751 metal Inorganic materials 0.000 description 5
- 239000002184 metal Substances 0.000 description 5
- 150000007530 organic bases Chemical class 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 239000003507 refrigerant Substances 0.000 description 5
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- YLEIFZAVNWDOBM-ZTNXSLBXSA-N ac1l9hc7 Chemical compound C([C@H]12)C[C@@H](C([C@@H](O)CC3)(C)C)[C@@]43C[C@@]14CC[C@@]1(C)[C@@]2(C)C[C@@H]2O[C@]3(O)[C@H](O)C(C)(C)O[C@@H]3[C@@H](C)[C@H]12 YLEIFZAVNWDOBM-ZTNXSLBXSA-N 0.000 description 4
- 239000007795 chemical reaction product Substances 0.000 description 4
- 229940126214 compound 3 Drugs 0.000 description 4
- 238000010511 deprotection reaction Methods 0.000 description 4
- 150000007524 organic acids Chemical class 0.000 description 4
- MFRIHAYPQRLWNB-UHFFFAOYSA-N sodium tert-butoxide Chemical compound [Na+].CC(C)(C)[O-] MFRIHAYPQRLWNB-UHFFFAOYSA-N 0.000 description 4
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 3
- QBXVXKRWOVBUDB-GRKNLSHJSA-N ClC=1C(=CC(=C(CN2[C@H](C[C@H](C2)O)C(=O)O)C1)OCC1=CC(=CC=C1)C#N)OCC1=C(C(=CC=C1)C1=CC2=C(OCCO2)C=C1)C Chemical compound ClC=1C(=CC(=C(CN2[C@H](C[C@H](C2)O)C(=O)O)C1)OCC1=CC(=CC=C1)C#N)OCC1=C(C(=CC=C1)C1=CC2=C(OCCO2)C=C1)C QBXVXKRWOVBUDB-GRKNLSHJSA-N 0.000 description 3
- 241000206595 Halomonas elongata Species 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- 241000801593 Pida Species 0.000 description 3
- LRIUKPUCKCECPT-UHFFFAOYSA-N [hydroxy(phenyl)-$l^{3}-iodanyl] 4-methylbenzenesulfonate Chemical compound C1=CC(C)=CC=C1S(=O)(=O)OI(O)C1=CC=CC=C1 LRIUKPUCKCECPT-UHFFFAOYSA-N 0.000 description 3
- 150000007513 acids Chemical class 0.000 description 3
- SRVFFFJZQVENJC-IHRRRGAJSA-N aloxistatin Chemical compound CCOC(=O)[C@H]1O[C@@H]1C(=O)N[C@@H](CC(C)C)C(=O)NCCC(C)C SRVFFFJZQVENJC-IHRRRGAJSA-N 0.000 description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 3
- 239000002585 base Substances 0.000 description 3
- SRSXLGNVWSONIS-UHFFFAOYSA-N benzenesulfonic acid Chemical compound OS(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-N 0.000 description 3
- 229940092714 benzenesulfonic acid Drugs 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- OSVHLUXLWQLPIY-KBAYOESNSA-N butyl 2-[(6aR,9R,10aR)-1-hydroxy-9-(hydroxymethyl)-6,6-dimethyl-6a,7,8,9,10,10a-hexahydrobenzo[c]chromen-3-yl]-2-methylpropanoate Chemical compound C(CCC)OC(C(C)(C)C1=CC(=C2[C@H]3[C@H](C(OC2=C1)(C)C)CC[C@H](C3)CO)O)=O OSVHLUXLWQLPIY-KBAYOESNSA-N 0.000 description 3
- 229940125782 compound 2 Drugs 0.000 description 3
- 229920002674 hyaluronan Polymers 0.000 description 3
- 229960003160 hyaluronic acid Drugs 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 229910000104 sodium hydride Inorganic materials 0.000 description 3
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 3
- PEZNEXFPRSOYPL-UHFFFAOYSA-N (bis(trifluoroacetoxy)iodo)benzene Chemical compound FC(F)(F)C(=O)OI(OC(=O)C(F)(F)F)C1=CC=CC=C1 PEZNEXFPRSOYPL-UHFFFAOYSA-N 0.000 description 2
- KBPLFHHGFOOTCA-UHFFFAOYSA-N 1-Octanol Chemical compound CCCCCCCCO KBPLFHHGFOOTCA-UHFFFAOYSA-N 0.000 description 2
- WGMHMVLZFAJNOT-UHFFFAOYSA-N 1-ethoxyethylideneazanium;chloride Chemical compound [Cl-].CCOC(C)=[NH2+] WGMHMVLZFAJNOT-UHFFFAOYSA-N 0.000 description 2
- BBMCTIGTTCKYKF-UHFFFAOYSA-N 1-heptanol Chemical compound CCCCCCCO BBMCTIGTTCKYKF-UHFFFAOYSA-N 0.000 description 2
- BLCJBICVQSYOIF-UHFFFAOYSA-N 2,2-diaminobutanoic acid Chemical compound CCC(N)(N)C(O)=O BLCJBICVQSYOIF-UHFFFAOYSA-N 0.000 description 2
- YIWUKEYIRIRTPP-UHFFFAOYSA-N 2-ethylhexan-1-ol Chemical compound CCCCC(CC)CO YIWUKEYIRIRTPP-UHFFFAOYSA-N 0.000 description 2
- WPHUUIODWRNJLO-UHFFFAOYSA-N 2-nitrobenzenesulfonyl chloride Chemical compound [O-][N+](=O)C1=CC=CC=C1S(Cl)(=O)=O WPHUUIODWRNJLO-UHFFFAOYSA-N 0.000 description 2
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 2
- DKPFZGUDAPQIHT-UHFFFAOYSA-N Butyl acetate Natural products CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
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- 206010013774 Dry eye Diseases 0.000 description 2
- 238000007167 Hofmann rearrangement reaction Methods 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 2
- AMQJEAYHLZJPGS-UHFFFAOYSA-N N-Pentanol Chemical compound CCCCCO AMQJEAYHLZJPGS-UHFFFAOYSA-N 0.000 description 2
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- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 description 2
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- 239000002537 cosmetic Substances 0.000 description 2
- ZSIAUFGUXNUGDI-UHFFFAOYSA-N hexan-1-ol Chemical compound CCCCCCO ZSIAUFGUXNUGDI-UHFFFAOYSA-N 0.000 description 2
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- 241000364051 Pima Species 0.000 description 1
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- OQWAXRPJEPTTSZ-UHFFFAOYSA-N [(2,3,4,5,6-pentafluorophenyl)-(2,2,2-trifluoroacetyl)oxy-$l^{3}-iodanyl] 2,2,2-trifluoroacetate Chemical compound FC1=C(F)C(F)=C(I(OC(=O)C(F)(F)F)OC(=O)C(F)(F)F)C(F)=C1F OQWAXRPJEPTTSZ-UHFFFAOYSA-N 0.000 description 1
- IBJMAUSPTVZWBK-UHFFFAOYSA-N [2-(2,2-dimethylpropanoyloxy)-3-iodophenyl] 2,2-dimethylpropanoate Chemical compound CC(C)(C)C(=O)OC1=CC=CC(I)=C1OC(=O)C(C)(C)C IBJMAUSPTVZWBK-UHFFFAOYSA-N 0.000 description 1
- RLRCQHZJIOEYAN-UHFFFAOYSA-N [acetyloxy-(2,3,4,5,6-pentafluorophenyl)-lambda3-iodanyl] acetate Chemical compound C1(F)=C(F)C(I(OC(=O)C)OC(=O)C)=C(F)C(F)=C1F RLRCQHZJIOEYAN-UHFFFAOYSA-N 0.000 description 1
- KCSMJUHAWHWVMR-UHFFFAOYSA-N [acetyloxy-(2-nitrophenyl)-$l^{3}-iodanyl] acetate Chemical compound CC(=O)OI(OC(C)=O)C1=CC=CC=C1[N+]([O-])=O KCSMJUHAWHWVMR-UHFFFAOYSA-N 0.000 description 1
- HHJIDOMMYDNFCA-UHFFFAOYSA-N [acetyloxy-(4-methylphenyl)-$l^{3}-iodanyl] acetate Chemical compound CC(=O)OI(OC(C)=O)C1=CC=C(C)C=C1 HHJIDOMMYDNFCA-UHFFFAOYSA-N 0.000 description 1
- SUYFTODEECUFSU-UHFFFAOYSA-N [methoxy(phenyl)-$l^{3}-iodanyl] 4-methylbenzenesulfonate Chemical compound C=1C=CC=CC=1I(OC)OS(=O)(=O)C1=CC=C(C)C=C1 SUYFTODEECUFSU-UHFFFAOYSA-N 0.000 description 1
- 102000005421 acetyltransferase Human genes 0.000 description 1
- 108020002494 acetyltransferase Proteins 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 150000001340 alkali metals Chemical class 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 150000001342 alkaline earth metals Chemical class 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 108010003977 aminoacylase I Proteins 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 239000000607 artificial tear Substances 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 229920001222 biopolymer Polymers 0.000 description 1
- 230000006696 biosynthetic metabolic pathway Effects 0.000 description 1
- SXDBWCPKPHAZSM-UHFFFAOYSA-N bromic acid Chemical compound OBr(=O)=O SXDBWCPKPHAZSM-UHFFFAOYSA-N 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 108010023417 cholesterol dehydrogenase Proteins 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- UKEFVSLHVRIWKJ-UHFFFAOYSA-N diethoxy(phenyl)-lambda3-iodane Chemical compound CCOI(OCC)C1=CC=CC=C1 UKEFVSLHVRIWKJ-UHFFFAOYSA-N 0.000 description 1
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 1
- SSTLWHMWDMWENH-UHFFFAOYSA-N dimethoxy(phenyl)-$l^{3}-iodane Chemical compound COI(OC)C1=CC=CC=C1 SSTLWHMWDMWENH-UHFFFAOYSA-N 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 238000000835 electrochemical detection Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012458 free base Substances 0.000 description 1
- ZZUFCTLCJUWOSV-UHFFFAOYSA-N furosemide Chemical compound C1=C(Cl)C(S(=O)(=O)N)=CC(C(O)=O)=C1NCC1=CC=CO1 ZZUFCTLCJUWOSV-UHFFFAOYSA-N 0.000 description 1
- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 150000004677 hydrates Chemical class 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 229910052747 lanthanoid Inorganic materials 0.000 description 1
- 150000002602 lanthanoids Chemical class 0.000 description 1
- JILPJDVXYVTZDQ-UHFFFAOYSA-N lithium methoxide Chemical compound [Li+].[O-]C JILPJDVXYVTZDQ-UHFFFAOYSA-N 0.000 description 1
- LZWQNOHZMQIFBX-UHFFFAOYSA-N lithium;2-methylpropan-2-olate Chemical compound [Li+].CC(C)(C)[O-] LZWQNOHZMQIFBX-UHFFFAOYSA-N 0.000 description 1
- AZVCGYPLLBEUNV-UHFFFAOYSA-N lithium;ethanolate Chemical compound [Li+].CC[O-] AZVCGYPLLBEUNV-UHFFFAOYSA-N 0.000 description 1
- 230000005923 long-lasting effect Effects 0.000 description 1
- 239000000395 magnesium oxide Substances 0.000 description 1
- CPLXHLVBOLITMK-UHFFFAOYSA-N magnesium oxide Inorganic materials [Mg]=O CPLXHLVBOLITMK-UHFFFAOYSA-N 0.000 description 1
- AXZKOIWUVFPNLO-UHFFFAOYSA-N magnesium;oxygen(2-) Chemical compound [O-2].[Mg+2] AXZKOIWUVFPNLO-UHFFFAOYSA-N 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229940073584 methylene chloride Drugs 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000001333 moisturizer Effects 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- LQNUZADURLCDLV-UHFFFAOYSA-N nitrobenzene Chemical compound [O-][N+](=O)C1=CC=CC=C1 LQNUZADURLCDLV-UHFFFAOYSA-N 0.000 description 1
- 238000010534 nucleophilic substitution reaction Methods 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 238000007867 post-reaction treatment Methods 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- LPNYRYFBWFDTMA-UHFFFAOYSA-N potassium tert-butoxide Chemical compound [K+].CC(C)(C)[O-] LPNYRYFBWFDTMA-UHFFFAOYSA-N 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- QDRKDTQENPPHOJ-UHFFFAOYSA-N sodium ethoxide Chemical compound [Na+].CC[O-] QDRKDTQENPPHOJ-UHFFFAOYSA-N 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 108010085346 steroid delta-isomerase Proteins 0.000 description 1
- 125000000472 sulfonyl group Chemical group *S(*)(=O)=O 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000004488 tear evaporation Effects 0.000 description 1
- 229910052723 transition metal Inorganic materials 0.000 description 1
- 150000003624 transition metals Chemical class 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
- C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
- C07D239/06—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
본 발명은 엑토인을 제조하는 방법에 관한 것으로서, L-글루타민에 노실(nosyl)기를 보호기로 도입하여 (2-노실)-L-글루타민을 제조하는 단계, 상기 화합물에 호프만 재배열 반응을 통해 (S)-4-아미노-(2-노실)부타노익산을 제조하는 단계, 노실기를 제거(탈보호화)하여 (S)-2,4-디아미노부타노익산을 제조하는 단계 및 유/무기 염기 존재 하에 알코올 용매에서 고리화반응을 진행하여 최종적으로 엑토인을 수득하는 단계를 포함하는 것을 특징으로 하는, 엑토인의 제조방법에 관한 것이다.The present invention relates to a method for producing ectoine, which includes introducing a nosyl group into L-glutamine to prepare (2-nosyl)-L-glutamine, and subjecting the compound to a Hoffman rearrangement reaction ( Preparing S)-4-amino-(2-nosyl)butanoic acid, removing (deprotecting) the nosyl group to prepare (S)-2,4-diaminobutanoic acid, and organic/inorganic It relates to a method for producing ectoine, which includes the step of performing a cyclization reaction in an alcohol solvent in the presence of a base to finally obtain ectoine.
Description
본 발명은 천연 사이클릭 아미노산인 엑토인(Ectoine)의 제조방법에 관한 것이다.The present invention relates to a method for producing ectoine, a natural cyclic amino acid.
엑토인(Ectoine)은 (S)-1,4,5,6-테트라하이드로-2-메틸-4-피리미딘카르복실산으로써, 여러 종의 박테리아에서 발견되는 천연 화합물이다. 극한 환경(예컨대 소금호수, 온천, 영구 빙설 등)에 사는 미생물로부터 얻어진 천연물질로서 극한 환경에서 스스로를 보호하기 위하여 천연물질인 엑토인을 형성한다. 엑토인의 분자구조는 하기 화학식 1에 의해 나타낼 수 있다.Ectoine is (S)-1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid, a natural compound found in several species of bacteria. It is a natural substance obtained from microorganisms living in extreme environments (e.g. salt lakes, hot springs, permanent ice and snow, etc.), and forms ectoin, a natural substance, to protect itself in extreme environments. The molecular structure of ectoine can be represented by the following formula (1).
[화학식 1][Formula 1]
엑토인은 친수성 미생물에서 고농도로 발견되며, 자연계에서 호염성 박테리아, 예컨대 할로모나스 엘롱가타(Halomonas elongata)를 통한 박테리아 과정을 통해서 자연적으로 생산되는 복소환 아미노산이다(E. A. Galinski et al., Eur. J. Biochem., 149(1985) pages 135-139.). Ectoine is a heterocyclic amino acid found in high concentrations in hydrophilic microorganisms and produced naturally through bacterial processes in nature through halophilic bacteria, such as Halomonas elongata (E. A. Galinski et al., Eur. J . Biochem., 149 (1985) pages 135-139.).
엑토인은 할로모나스 엘롱가타로 불리우는 호열성 박테리아로부터 추출될 수 있고, 여기서 엑토인은 박테리아에서 삼투압을 보상하고 생체중합체를 안정화시켜 탈수, 고온 및 UV 손상으로부터 박테리아를 보호한다.Ectoine can be extracted from a thermophilic bacterium called Halomonas elongata, where ectoine compensates for osmotic pressure in the bacteria and stabilizes the biopolymer, protecting the bacteria from dehydration, high temperatures and UV damage.
미국특허 US08/953,988에 의하면, 엑토인은 보습력이 탁월하며, 항산화제(antioxidant)나 방부제(preservative) 없이도 매우 높은 안정성을 유지하며, 화장품용으로 사용 가능한 제품이며 머크(Merck)사에서 로나케어 히드로인(RonaCare® hydoine) 제품이 판매되고 있는 실정이다.According to U.S. Patent US08/953,988, ectoin has excellent moisturizing properties and maintains very high stability even without antioxidants or preservatives. It is a product that can be used for cosmetics and is sold as Ronacare Hydro by Merck. Phosphorus (RonaCare® hydoine) products are being sold.
그 외에도 미세먼지의 영향에 의해 유발된 폐질환의 치료가 WO2005/002556에 기술되어 있고, 바이오센서에서의 엑토인에 의한 글루코오스 옥시다아제의 안정화가 WO2007/097653에 기재되어 있으며, 효소 활성의 전기화학적 검출을 목적으로 콜레스테롤 디히드로게나아제를 포함하는 콜레스테롤 바이오센서에서의 시약으로 WO2007/132226에서 보고된 바 있다.In addition, treatment of lung diseases caused by the effects of fine dust is described in WO2005/002556, stabilization of glucose oxidase by ectoin in biosensors is described in WO2007/097653, and electrochemical detection of enzyme activity For this purpose, it has been reported in WO2007/132226 as a reagent in a cholesterol biosensor containing cholesterol dehydrogenase.
최근에는 안과학적으로 사용 가능함이 밝혀지고 있는데, 유럽특허 EP0671161에서는 피부의 수분함량을 증가시키는 화장품에서 보습제로 사용될 수 있는 것을 기재하고 있고, DE10-2014-007423에서는 엑토인을 함유하는 눈의 염증 치료용 조성물이 개시되어 있다. 이 특허에서는 건성각 결막염(Kerotoconjunctivities sicca)을 치료하는 것이 히알루론산(Hyaluronic acid) 용액으로 치료하는 것보다 효과적이라는 것이 밝혀짐으로써 기존의 인공눈물로 사용되었던 히알루론산과 더불어 엑토인의 사용범위가 더욱 넓어질 수 있다는 것을 의미한다.Recently, it has been revealed that it can be used ophthalmologically. European patent EP0671161 describes that it can be used as a moisturizer in cosmetics that increases the moisture content of the skin, and DE10-2014-007423 describes treatment of eye inflammation containing ectoin. A composition for use is disclosed. In this patent, it was revealed that treating keratoconjunctivitis sicca is more effective than treating hyaluronic acid solution, expanding the scope of use of ectoin along with hyaluronic acid, which was previously used as artificial tears. This means that it can be expanded.
히알루론산과 엑토인을 같이 사용하면, 눈물의 증발로부터의 보호가 오래 지속되며, 안구 건조증, 즉 건조 증후군(sicca syndrome)의 치료 또는 예방, 결막염(conjunctivitis) 및 고초열(hayfever)의 치료 및 예방에 적합한 것으로 보고되고 있다.When used together, hyaluronic acid and ectoine provide long-lasting protection against tear evaporation and are used in the treatment or prevention of dry eye syndrome (sicca syndrome), treatment and prevention of conjunctivitis and hayfever. It is reported to be suitable for .
이러한 엑토인의 제조방법을 살펴보면, 현재는 주로 생합성경로(Biosynthetic pathway)로 생산하고 있으며, 독일의 비톱(Bitop AG)이 유일하다. 엑토인의 생합성은 디아미노부타이릭산(DABA) 아미노트랜스퍼라제(EctB), 아세틸트랜스퍼라제(EctA) 및 엑토인 신터세이즈(EctC)를 사용하는 방법이며, 하기 반응식 1에 구체적인 반응경로를 도시하였다.Looking at the manufacturing method of ectoine, it is currently mainly produced through the biosynthetic pathway, and Germany's Bitop AG is the only one. The biosynthesis of ectoine uses diaminobutyric acid (DABA) aminotransferase (EctB), acetyltransferase (EctA), and ectoine sinterase (EctC), and the specific reaction pathway is shown in Scheme 1 below. .
[반응식 1][Scheme 1]
Bitop사가 개발한 방법으로써, 바이오밀킹(Biomilking)이라고도 불리우며, 생산품 용액을 전기영동과 크로마토그래피, 여과 후 농축 결정화를 진행함으로써 수득하며, 최근에는 H. elongata의 연속적인 발효공정을 이용하여 대량생산 방법을 개발하였다고 보고하고 있다. 상기 공정을 영원한 밀킹(permanent milking) 프로세스라고도 칭하고 있다.It is a method developed by Bitop, also called biomilking, and is obtained by performing electrophoresis, chromatography, filtration, and concentration and crystallization of the product solution. Recently, a mass production method has been developed using the continuous fermentation process of H. elongata. It is reported that it has been developed. The above process is also called a permanent milking process.
또한, Bitop의 미국특허 US8,598,346을 보면, 아미노아실레이즈를 이용한 방법도 개시하고 있으나, 합성공정이 길어서 엑토인의 제조원가와 경제성 면에서는 비효율적인 방법이라고 사료된다. 제조방법은 하기 반응식 2에 도시하였다.In addition, Bitop's U.S. patent US8,598,346 discloses a method using aminoacylase, but it is considered an inefficient method in terms of manufacturing cost and economic feasibility of ectoine due to the long synthesis process. The manufacturing method is shown in Scheme 2 below.
[반응식 2][Scheme 2]
중국특허 CN108003105에서는 아세틸-L-글루타민을 이용하여 엑토인을 합성하는 방법을 개시하고 있는데, 대량생산에는 적합하지 않은 디사이클로헥산카보디이미드(DCC), 소듐하이드라이드(NaH), 트리에틸실란(Et3SiH)을 사용하는 공정이어서 소량으로는 제조 가능하나 대량생산에는 적합하지 않은 방법이다. 이 제조방법은 하기 반응식 3에 도시하였다.Chinese patent CN108003105 discloses a method of synthesizing ectoine using acetyl-L-glutamine, but dicyclohexanecarbodiimide (DCC), sodium hydride (NaH), and triethylsilane ( Since it is a process using Et 3 SiH), it can be manufactured in small quantities, but is not suitable for mass production. This manufacturing method is shown in Scheme 3 below.
[반응식 3][Scheme 3]
상술한 바와 같은 문제점을 해결하고자, 본 발명의 발명자는 경제적이면서 대량생산에 적합한 방법을 강구하였으며, 마침내 새로운 제조방법을 개발하게 된 것이다.In order to solve the problems described above, the inventor of the present invention sought a method that was economical and suitable for mass production, and eventually developed a new manufacturing method.
(특허문헌 1) JP 03031265 A (Takeda Chem Ind Ltd), 1989/06/26(Patent Document 1) JP 03031265 A (Takeda Chem Ind Ltd), 1989/06/26
(특허문헌 2) US 0953988 A (Merck Patent GmbH), 1993/12/14(Patent Document 2) US 0953988 A (Merck Patent GmbH), 1993/12/14
(특허문헌 3) US 8598346 B2 (Bitop AG), 2008/07/16(Patent Document 3) US 8598346 B2 (Bitop AG), 2008/07/16
(비특허문헌 1) E. A. Galinski et al., Eur. J. Biochem.m 149(1985) pages 135-139.(Non-patent Document 1) E. A. Galinski et al., Eur. J. Biochem. m 149 (1985) pages 135-139.
본 발명은 상기와 같은 문제점을 해결하기 위해 안출된 것으로,The present invention was devised to solve the above problems,
본 발명은 고압 반응기와 같은 고가의 제조설비가 불필요하며, 반응 및 반응 후의 처리가 간편하고 효율적으로 엑토인 화합물을 제조할 수 있는 제조방법을 제공하는데 그 목적이 있다.The purpose of the present invention is to provide a production method that does not require expensive production equipment such as a high-pressure reactor and can produce ectoine compounds easily and efficiently in reaction and post-reaction treatment.
한편, 본 발명에 있어 명시되지 않은 또 다른 목적들은 하기의 과제 해결 수단, 발명의 효과 및 상세한 설명으로부터 용이하게 추론 가능한 범위 내에서 추가적으로 고려될 것이다.Meanwhile, other purposes not specified in the present invention will be additionally considered within the scope that can be easily inferred from the problem solving means, effects of the invention, and detailed description below.
상술한 바와 같은 해결과제를 달성하기 위한 엑토인의 제조방법은, L-글루타민과 나이트로(Nitro) 치환된 벤젠 설포닐 클로라이드를 반응시켜 2-노실(nosyl) 치환된 L-글루타민을 제조하고 이어서 노실(nosyl) 보호화된 L-글루타민에 호프만 재배열반응(hofmann rearrangement reaction)을 진행한 후 탈보호화반응을 통하여 (S)-2,4-디아미노 부타노익산을 제조한 후 고리화 반응을 수행하여 최종적으로 엑토인을 제조하는 방법이다.The method for producing ectoine to achieve the above-described problem is to react L-glutamine with nitro-substituted benzene sulfonyl chloride to produce 2-nosyl-substituted L-glutamine, followed by After performing a Hofmann rearrangement reaction on nosyl-protected L-glutamine, (S)-2,4-diaminobutanoic acid was prepared through a deprotection reaction, followed by a cyclization reaction. This is the final method of producing ectoin.
따라서, 본 발명의 엑토인의 제조방법은 다음과 같은 구성을 가진 실시예에 의해서 구현된다.Accordingly, the method for producing ectoine of the present invention is implemented by an example having the following configuration.
본 발명의 일 실시예에 따르면, 본 발명에 따른 엑토인의 제조방법은, (ⅰ) L-글루타민 및 나이트로-치환 벤젠설포닐 클로라이드를 포함하는 혼합물을 염기조건 하에서 반응시키는 과정을 포함하는 공정을 통해 하기 화학식 1로 표시되는 제1중간체를 수득하는 제1단계;According to one embodiment of the present invention, the method for producing ectoine according to the present invention includes (i) reacting a mixture containing L-glutamine and nitro-substituted benzenesulfonyl chloride under basic conditions. A first step of obtaining a first intermediate represented by the following formula (1);
[화학식 1][Formula 1]
(상기 화학식 1에서, R1은 NO2 또는 H이고, R2는 NO2 또는 H이다)(In Formula 1, R1 is NO 2 or H, and R2 is NO 2 or H)
(ⅱ) 상기 제1단계에서 수득한 상기 제1중간체 및 하이퍼발란트 아이오딘 시약(hypervalent iodine reagent)을 포함하는 혼합물을 반응시키는 과정을 포함하는 공정을 통해 하기 화학식 2로 표시되는 제2중간체를 수득하는 제2단계;(ii) producing a second intermediate represented by the following formula (2) through a process comprising reacting a mixture containing the first intermediate and a hypervalent iodine reagent obtained in the first step. The second step of obtaining;
[화학식 2][Formula 2]
(상기 화학식 1에서, R1은 NO2 또는 H이고, R2는 NO2 또는 H이다)(In Formula 1, R1 is NO 2 or H, and R2 is NO 2 or H)
(ⅲ) 상기 제2단계에서 수득한 상기 제2중간체 및 티올계 화합물을 포함하는 혼합물을 염기조건 하의 유기용매에 투입, 반응시키는 과정을 포함하는 공정을 통해 하기 화학식 3으로 표시되는 (S)-2,4-디아미노부타노익산을 수득하는 제3단계;(iii) Through a process including adding the mixture containing the second intermediate and the thiol-based compound obtained in the second step to reacting in an organic solvent under basic conditions, (S)- represented by the following formula 3 is obtained. The third step of obtaining 2,4-diaminobutanoic acid;
[화학식 3][Formula 3]
(ⅳ) 상기 제3단계에서 수득한 상기 (S)-2,4-디아미노부타노익산, 알킬아세트이미데이트 화합물 및 염기를 포함하는 혼합물을 유기용매에 투입, 반응시키는 과정을 포함하는 공정을 통해 하기 화학식 4로 표시되는 엑토인(Ectoine)을 수득하는 제4단계;를 포함한다.(iv) a process comprising adding the mixture containing (S)-2,4-diaminobutanoic acid, alkylacetimidate compound, and base obtained in the third step to an organic solvent and reacting it. It includes a fourth step of obtaining ectoine represented by the following formula (4).
[화학식 4][Formula 4]
본 발명의 다른 실시예에 따르면, 본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제2단계의 하이퍼발란트 아이오딘 시약은 하기 화학식 5로 표시되는 화합물인 것을 특징으로 한다.According to another embodiment of the present invention, in the method for producing ectoine according to the present invention, the hypervalent iodine reagent in the second step is a compound represented by the following formula (5).
[화학식 5][Formula 5]
본 발명의 또 다른 실시예에 따르면, 본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제2단계의 혼합물은 상기 하이퍼발란트 아이오딘 시약이 상기 제1중간체 대비 1.0 내지 3.0 당량비로 포함되어 있는 혼합물인 것을 특징으로 한다.According to another embodiment of the present invention, in the method for producing ectoine according to the present invention, the mixture in the second step contains the hypervalent iodine reagent in an equivalent ratio of 1.0 to 3.0 compared to the first intermediate. It is characterized as a mixture.
본 발명의 또 다른 실시예에 따르면, 본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제3단계의 티올계 화합물은 알킬티올, 방향족티올 또는 티오글리콜산인 것을 특징으로 하며, 상기 알킬티올의 알길기는 탄소수 1~8의 직쇄형 알킬기 중 어느 하나인 것을 특징으로 한다.According to another embodiment of the present invention, in the method for producing ectoine according to the present invention, the thiol-based compound in the third step is an alkylthiol, aromatic thiol, or thioglycolic acid, and the alkylthiol's alkylthiol The long group is characterized as being any one of a straight-chain alkyl group having 1 to 8 carbon atoms.
본 발명의 또 다른 실시예에 따르면, 본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제4단계의 혼합물은 상기 염기가 상기 (S)-2,4-디아미노부타노익산 대비 1.0 내지 3.0 당량비로 포함되어 있는 혼합물인 것을 특징으로 한다.According to another embodiment of the present invention, in the method for producing ectoine according to the present invention, the mixture in the fourth step has a base content of 1.0 to 3.0 relative to the (S)-2,4-diaminobutanoic acid. It is characterized as a mixture contained in an equivalent ratio.
본 발명은 상기 개시한 과제의 해결 수단을 채용함으로써 하기와 같은 효과를 가진다.The present invention has the following effects by employing means for solving the problems disclosed above.
본 발명의 제조방법은 제조공정 중에서 발생하는 반응부산물을 별도의 정제과정 없이, 동일 제조 공정에서 효과적으로 정제(또는 제거)할 수 있고, 고압이나 고온의 반응조건이 불필요하며, 단순한 제조공정으로 높은 수율 및 높은 순도의 엑토인을 높은 생산성으로 제조할 수 있는 효과가 있다.The manufacturing method of the present invention can effectively purify (or remove) reaction by-products generated during the manufacturing process in the same manufacturing process without a separate purification process, does not require high pressure or high temperature reaction conditions, and provides high yield with a simple manufacturing process. And it has the effect of producing high purity ectoine with high productivity.
한편, 본 발명에 있어 명시적으로 언급되지 않은 효과들이라도 하기의 상세한 설명 등 명세서 전체의 기재에 있어 합리적으로 추론 가능한 범위 내에서 도출 가능한 효과라면 본 명세서에 기재된 것으로 취급할 수 있음은 물론이다. Meanwhile, it goes without saying that even if effects are not explicitly mentioned in the present invention, if they are effects that can be derived within a range that can be reasonably inferred from the description of the entire specification, such as the detailed description below, they can be treated as described in the present specification.
도 1은 본 발명에서의 실시예에 따라 수득되는 제1중간체에 해당하는 화합물 2a의 1H NMR
도 2는 본 발명에서의 실시예에 따라 수득되는 제1중간체에 해당하는 화합물 2a의 13C NMR
도 3은 본 발명에서의 실시예에 따라 수득되는 제1중간체에 해당하는 화합물 2b의 1H NMR
도 4는 본 발명에서의 실시예에 따라 수득되는 제1중간체에 해당하는 화합물 2b의 13C NMR
도 5는 본 발명에서의 실시예에 따라 수득되는 제1중간체에 해당하는 화합물 2c의 1H NMR
도 6은 본 발명에서의 실시예에 따라 수득되는 제1중간체에 해당하는 화합물 2c의 13C NMR
도 7은 본 발명에서의 실시예에 따라 수득되는 제2중간체에 해당하는 화합물 3a의 1H NMR
도 8은 본 발명에서의 실시예에 따라 수득되는 제2중간체에 해당하는 화합물 3a의 13C NMR
도 9는 본 발명에서의 실시예에 따라 수득되는 제2중간체에 해당하는 화합물 3b의 1H NMR
도 10은 본 발명에서의 실시예에 따라 수득되는 제2중간체에 해당하는 화합물 3b의 13C NMR
도 11은 본 발명에서의 실시예에 따라 수득되는 제2중간체에 해당하는 화합물 3c의 1H NMR
도 12는 본 발명에서의 실시예에 따라 수득되는 제2중간체에 해당하는 화합물 3c의 13C NMR
도 13은 본 발명에서의 실시예에 따라 수득되는 (S)-2,4-디아미노부타노익산의 1H NMR
도 14는 본 발명에서의 실시예에 따라 수득되는 (S)-2,4-디아미노부타노익산의 13C NMR
도 15는 본 발명에서의 실시예에 따라 수득되는 엑토인의 1H NMR
도 16은 본 발명에서의 실시예에 따라 수득되는 엑토인의 13C NMR1 shows 1 H NMR of compound 2a, which corresponds to the first intermediate obtained according to an example of the present invention.
Figure 2 is 13 C NMR of compound 2a corresponding to the first intermediate obtained according to the examples of the present invention.
Figure 3 shows 1 H NMR of compound 2b, which corresponds to the first intermediate obtained according to the examples of the present invention.
Figure 4 shows 13 C NMR of compound 2b, which corresponds to the first intermediate obtained according to the examples of the present invention.
Figure 5 is 1 H NMR of compound 2c, which corresponds to the first intermediate obtained according to the examples of the present invention.
Figure 6 shows 13 C NMR of compound 2c, which corresponds to the first intermediate obtained according to the examples of the present invention.
Figure 7 shows 1 H NMR of compound 3a, which corresponds to the second intermediate obtained according to the examples of the present invention.
Figure 8 shows 13 C NMR of compound 3a, which corresponds to the second intermediate obtained according to the examples of the present invention.
Figure 9 shows 1 H NMR of compound 3b, which corresponds to the second intermediate obtained according to the examples of the present invention.
Figure 10 shows 13 C NMR of compound 3b, which corresponds to the second intermediate obtained according to the examples of the present invention.
Figure 11 shows 1 H NMR of compound 3c, which corresponds to the second intermediate obtained according to the examples of the present invention.
Figure 12 shows 13 C NMR of compound 3c, which corresponds to the second intermediate obtained according to the examples of the present invention.
13 is 1 H NMR of (S)-2,4-diaminobutanoic acid obtained according to an example of the present invention.
Figure 14 shows 13 C NMR of (S)-2,4-diaminobutanoic acid obtained according to the examples of the present invention.
15 is 1 H NMR of ectoine obtained according to an example in the present invention.
Figure 16 shows 13 C NMR of ectoine obtained according to the examples in the present invention.
이하에서는 본 발명에 따른 엑토인의 제조방법의 바람직한 실시예들을 첨부된 도면을 참조하여 상세히 설명한다. 하기에서 본 발명을 설명함에 있어서 공지 기능 또는 구성에 대한 구체적인 설명이 본 발명의 요지를 불필요하게 흐릴 수 있다고 판단되는 경우에는 그 상세한 설명을 생략하도록 한다. 명세서 전체에서, 어떤 부분이 어떤 구성요소를 "포함"한다고 할 때 이는 특별히 반대되는 기재가 없는 한 다른 구성요소를 제외하는 것이 아니라 다른 구성요소를 더 포함할 수 있는 것을 의미한다.Hereinafter, preferred embodiments of the method for producing ectoine according to the present invention will be described in detail with reference to the attached drawings. In the following description of the present invention, if a detailed description of a known function or configuration is judged to unnecessarily obscure the gist of the present invention, the detailed description will be omitted. Throughout the specification, when a part "includes" a certain component, this means that it may further include other components rather than excluding other components, unless specifically stated to the contrary.
다른 정의가 없다면, 본 명세서에서 사용되는 모든 용어(기술 및 과학적 용어를 포함)는 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자에게 공통적으로 이해될 수 있는 의미로 사용될 수 있을 것이다. 또 일반적으로 사용되는 사전에 정의되어 있는 용어들은 명백하게 특별히 정의되지 않는 한 이상적으로 또는 과도하게 해석되지 않는다.Unless otherwise defined, all terms (including technical and scientific terms) used in this specification may be used with meanings that can be commonly understood by those skilled in the art to which the present invention pertains. Additionally, terms defined in commonly used dictionaries are not interpreted ideally or excessively unless clearly specifically defined.
본 발명의 일 실시예에 따른 엑토인의 제조방법은, L-글루타민과 나이트로(nitro)-치환 벤젠설포닐 클로라이드를 반응시켜 제1중간체인 2-노실(nosyl)-L-글루타민을 제조하는 제1단계; 상기 2-노실-L-글루타민에 하이퍼발란트 아이오딘 시약을 투입하여 호프만 재배열 반응(Hofmann rearrangement reaction)을 유도하여 제2중간체인 (S)-4-아미노-(2-노실)부타노익산을 제조하는 제2단계; 상기 (S)-4-아미노-(2-노실)부타노익산을 염기 조건 하에 알킬티올 또는 방향족 티올 등 티올계 화합물을 첨가, 탈보호화하여 (S)-2,4-디아미노 부타노익산을 제조하는 제3단계; 상기 (S)-2,4-디아미노 부타노익산에 트리알킬오쏘아세테이트(trialkylorthoacetate)(또는 알킬아세트이미데이트 화합물), 무기/유기 염기를 사용하고 알코올계 유기용매 하에서 고리화반응을 수행하여 엑토인을 제조하는 제4단계;를 포함한다.The method for producing ectoine according to an embodiment of the present invention involves reacting L-glutamine with nitro-substituted benzenesulfonyl chloride to produce 2-nosyl-L-glutamine, the first intermediate. Step 1; Hypervalent iodine reagent is added to the 2-nosyl-L-glutamine to induce a Hofmann rearrangement reaction to produce the second intermediate, (S)-4-amino-(2-nosyl)butanoic acid. The second step of manufacturing; The (S)-4-amino-(2-nosyl)butanoic acid is deprotected by adding a thiol-based compound such as alkylthiol or aromatic thiol under basic conditions to produce (S)-2,4-diaminobutanoic acid. Third stage of manufacturing; Trialkylorthoacetate (or alkylacetimidate compound) and an inorganic/organic base are used in the (S)-2,4-diaminobutanoic acid, and a cyclization reaction is performed in an alcohol-based organic solvent to obtain ecto. It includes a fourth step of producing phosphorus.
본 발명에서는, 글루타민의 아민기를 보호하기 위해 노실(나이트로벤젠설포닐)기를 사용하였는데, 이러한 노실 보호기를 사용한 것은 탈보호화반응 시 티올의 방향족 친핵성 치환반응이 쉽게 일어나면서 마이젠하이머 복합체(Meisenheimer complex)가 중간체로 잘 형성되며, 염기조건 하에서 쉽게 탈보호화가 되기에 이점을 가질 수 있기 때문이다. 탈보호화반응을 산조건하에서 수행하면 원하지 않는 부산물이 많이 생성되어 불순물을 제거하는데 애로사항이 생길 수 있다.In the present invention, a nosyl (nitrobenzenesulfonyl) group was used to protect the amine group of glutamine. The use of this nosyl protecting group facilitates the aromatic nucleophilic substitution reaction of thiol during the deprotection reaction, forming the Meisenheimer complex. ) is easily formed as an intermediate and can be easily deprotected under basic conditions, which can be advantageous. If the deprotection reaction is performed under acid conditions, many unwanted by-products may be generated, which may cause difficulties in removing impurities.
본 발명에서의 각 단계를 보다 구체적으로 설명하면, 상기 제1단계의 경우 L-글루타민, 나이트로-치환 벤젠설포닐 클로라이드(노실(바람직하게는 2-나이트로벤젠설포닐, 4-나이트로벤젠설포닐, 2-4-다이나이트로벤젠설포닐 중 어느 하나) 클로라이드), 정제수를 차례대로 투입하고 염기조건 하에서 내부온도 25℃ 이하에서 2시간 동안 교반하여 결정화하는 과정을 의미할 수 있고, 상기 제2단계의 경우 상기 제1단계에서 수득한 제1중간체인 2-노실-L-글루타민과 아세토나이트릴, 에틸아세테이트, 증류수를 차례대로 투입하여 완전히 용해시키고 내부온도를 5℃ 이하로 냉각시킨 다음, 하이퍼발렌트 아이오딘 시약(예컨대 (다이아세톡시아이오도)벤젠(PIDA))을 3회 소분하여 30분 동안 반응액에 투입하고 내부온도 0 ~ 5℃에서 5시간 동안 교반한 뒤 추출, 농축, 결정화시킴으로써 호프만 재배열 반응을 일으키는 과정을 의미할 수 있으며, 상기 제3단계의 경우 상기 제2단계에서 수득한 제2중간체인 (S)-4-아미노-(2-노실)부타노익산, 티올계 화합물(예컨대 티오페놀), 포타슘 하이드록사이드, 용매(예컨대 아세토니트릴) 및 증류수를 투입하고 2시간 동안 55℃에서 교반한 뒤 농축, 결정화시키는 탈보호화 반응을 일으키는 과정을 의미할 수 있으며, 상기 제4단계의 경우 상기 제3단계에서 수득한 (S)-2,4-디아미노부타노익산, 알코올계 용매(예컨대 n-부탄올), 알킬아세트이미데이트 염산(예컨대 에틸아세트이미데이트 염산), 유기 염기(예컨대 트리에틸아민), 무기염기(예컨대 소듐 t-부톡사이드)를 투입하고 24시간 동안 환류교반한 뒤 농축, 결정화함으로써 고리화반응을 일으키는 과정을 의미할 수 있다.To describe each step in the present invention in more detail, in the case of the first step, L-glutamine, nitro-substituted benzenesulfonyl chloride (nosyl (preferably 2-nitrobenzenesulfonyl, 4-nitrobenzene) It may refer to the process of crystallization by sequentially adding sulfonyl, 2-4-dinitrobenzenesulfonyl (chloride), and purified water and stirring for 2 hours at an internal temperature of 25°C or lower under basic conditions, In the second step, 2-nosyl-L-glutamine, the first intermediate obtained in the first step, acetonitrile, ethyl acetate, and distilled water are added in order to completely dissolve and the internal temperature is cooled to 5°C or lower. , hypervalent iodine reagent (e.g. (diacetoxyiodo)benzene (PIDA)) was divided into three portions and added to the reaction solution for 30 minutes, stirred for 5 hours at an internal temperature of 0 to 5°C, then extracted and concentrated. , can refer to the process of causing a Hoffmann rearrangement reaction by crystallizing, and in the case of the third step, (S)-4-amino-(2-nosyl)butanoic acid, the second intermediate obtained in the second step, It may refer to the process of adding a thiol-based compound (e.g. thiophenol), potassium hydroxide, solvent (e.g. acetonitrile), and distilled water, stirring at 55°C for 2 hours, and then causing a deprotection reaction of concentration and crystallization. In the case of the fourth step, (S)-2,4-diaminobutanoic acid obtained in the third step, an alcoholic solvent (e.g. n-butanol), alkylacetimidate hydrochloric acid (e.g. ethylacetimidate hydrochloric acid) , may refer to a process of causing a cyclization reaction by adding an organic base (e.g., triethylamine) and an inorganic base (e.g., sodium t-butoxide), refluxing and stirring for 24 hours, and then concentrating and crystallizing.
종합적으로, 하기 반응식 4에서 도시하는 바와 같이 본 발명은 엑토인 합성의 핵심중간체인 2-노실-L-글루타민(하기 반응식 4에서의 화합물 2)을 제조하고, 호프만 재배열 반응을 통해 상기 2-노실-L-글루타민으로부터 (S)-4-아미노-(2-노실)부타노익산(하기 반응식 4에서의 화합물 3)을 제조 후, 상기 화합물 3을 탈보호화한 뒤 고리화하여 엑토인을 제조하는 방법에 관한 것이다.Overall, as shown in Scheme 4 below, the present invention prepares 2-nosyl-L-glutamine (compound 2 in Scheme 4 below), a key intermediate in ectoine synthesis, and produces the 2- through the Hoffman rearrangement reaction. After preparing (S)-4-amino-(2-nosyl)butanoic acid (compound 3 in Scheme 4 below) from nosyl-L-glutamine, ectoine was prepared by deprotecting the compound 3 and then cyclizing it. It's about how to do it.
[반응식 4][Scheme 4]
상기 반응식 4에 있어 핵심중간체 화합물인 화합물 2(2a, 2b, 2c) 및 화합물 3(3a, 3b, 3c)을 순수하게 분리하고 그 구조를 규명하고자 하는 시도는 지금까지 없었다. 따라서, 본 발명에서 신규하게 합성한 상기 화합물들에 대한 구조 규명을 수행하였고, 이에 대한 데이터는 후술하기로 한다. 상술한 바와 같이 화합물 2(2a, 2b, 2c)는 상기 제1중간체에 해당하며, 화합물 3(3a, 3b, 3c)는 상기 제2중간체에 해당한다.There has been no attempt so far to purely isolate Compound 2 (2a, 2b, 2c) and Compound 3 (3a, 3b, 3c), which are the key intermediate compounds in Scheme 4, and identify their structures. Therefore, the structures of the compounds newly synthesized in the present invention were investigated, and data on this will be described later. As described above, compound 2 (2a, 2b, 2c) corresponds to the first intermediate, and compound 3 (3a, 3b, 3c) corresponds to the second intermediate.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제2단계에 있어 하이퍼발란트 아이오딘 시약은, 하기 [화학식 2]로 표현되는 화합물을 의미할 수 있다.In the method for producing ectoine according to the present invention, the hypervalent iodine reagent in the second step may refer to a compound represented by the following [Formula 2].
[화학식 2][Formula 2]
이때 상기 화합물들로는, (다이아세톡시아이오도)벤젠(PIDA), 비스(트리플루오로아세톡시)아이오도벤젠(PIFA), 비스(t-부틸카보닐옥시)아이오도벤젠(PhI(Piv)2), 아세트산 납(Lead Ⅳ acetate) 또는 코져시약(Koser's reagent), 즉 [하이드록시(토실옥시)아이오도]벤젠(HTIB), [메톡시(토실옥시)아이오도]벤젠(MTIB), 4-(디아세톡시아이오도)톨루엔, 2-나이트로-(다이아세톡시아이오도)벤젠, 2-나이트로-(비스(트리플루오로아세톡시)아이오도)벤젠, (다이아섹톡시아이오도)펜타플루오로벤젠(C6F5I(OAc)2), 비스(트리플루오로아세톡시)아이오도펜타플루오로벤젠(C6F5I(CF3)2), (다이메톡시아이오도)벤젠(PIMA) 및 (다이에톡시아이오도)벤젠(PIEA)으로 이루어진 군으로부터 선택된 1종 이상을 의미할 수 있다.At this time, the compounds include (diacetoxyiodo)benzene (PIDA), bis(trifluoroacetoxy)iodobenzene (PIFA), and bis(t-butylcarbonyloxy)iodobenzene (PhI(Piv) 2 ), Lead Ⅳ acetate or Koser's reagent, i.e. [hydroxy(tosyloxy)iodo]benzene (HTIB), [methoxy(tosyloxy)iodo]benzene (MTIB), 4- (diacetoxyiodo)toluene, 2-nitro-(diacetoxyiodo)benzene, 2-nitro-(bis(trifluoroacetoxy)iodo)benzene, (diacetoxyiodo)penta Fluorobenzene (C 6 F 5 I(OAc) 2 ), bis(trifluoroacetoxy)iodopentafluorobenzene (C 6 F 5 I(CF 3 ) 2 ), (dimethoxyiodo)benzene It may refer to one or more types selected from the group consisting of (PIMA) and (diethoxyiodo)benzene (PIEA).
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제2단계는 용매 하에서 수행될 수 있으며, 상기 유기용매는 아세토니트릴, 테트라하이드로퓨란(THF), 디메틸포름아마이드, 디메틸설폭사이드, 디메틸아세트아마이드, 아세톤, 에틸아세테이트, 메틸아세테이트, 아이소프로필아세테이트, 부틸아세테이트, 메틸렌클로라이드 및 N-메틸피롤리돈으로 이루어진 군으로부터 선택된 1종 이상 또는 이들의 혼합물을 의미할 수 있다.In the method for producing ectoine according to the present invention, the second step may be performed in a solvent, and the organic solvent is acetonitrile, tetrahydrofuran (THF), dimethylformamide, dimethyl sulfoxide, dimethylacetamide, It may mean one or more selected from the group consisting of acetone, ethyl acetate, methyl acetate, isopropyl acetate, butylacetate, methylene chloride, and N-methylpyrrolidone, or a mixture thereof.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제2단계의 혼합 및 반응은 출발물질인 상기 L-글루타민 1 당량에 대하여 상기 하이퍼발란트 아이오딘 시약을 0.9 내지 3.0 당량비, 바람직하게는 1.0 내지 2.0 당량비, 더욱 바람직하게는 1.0 내지 1.4 당량비로 혼합하여 수행할 수 있다. 상기 하이퍼발란트 아이오딘 시약의 당량비가 0.9 미만이면 목적하는 반응 생성물 수율이 낮은 문제가 발생할 수 있고, 3.0 당량비를 초과하면 과사용된 하이퍼발란트 아이오딘 시약이 불순물로 작용하여 목적하는 최종 반응생성물의 순도가 떨어지며, 제조단가가 크게 상승하는 문제가 있다.In the method for producing ectoine according to the present invention, the mixing and reaction in the second step is performed by using the hypervalent iodine reagent in an equivalent ratio of 0.9 to 3.0, preferably 1.0 to 1.0, relative to 1 equivalent of L-glutamine, which is the starting material. It can be performed by mixing at an equivalent ratio of 2.0, more preferably at an equivalent ratio of 1.0 to 1.4. If the equivalence ratio of the hypervalent iodine reagent is less than 0.9, a problem may occur in which the yield of the desired reaction product is low, and if it exceeds 3.0, the overused hypervalent iodine reagent may act as an impurity and reduce the desired final reaction product. There is a problem that the purity is low and the manufacturing cost increases significantly.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제2단계는 -20 내지 40℃의 온도에서 1 내지 24시간 동안, 바람직하게는 -10 내지 30℃의 온도에서 1 내지 12시간 동안, 더욱 바람직하게는 0 내지 10℃의 온도에서 1 내지 6시간 동안 교반시키면서 수행하는 것을 특징으로 할 수 있다. 상기 반응온도가 -10℃ 미만이면 반응온도가 너무 낮아 교반이 어려울 수 있으며, 반응온도가 40℃를 초과하면 고리화 반응이 진행되어 불순물이 증가하는 결과가 발생하는 문제가 있다.In the method for producing ectoine according to the present invention, the second step is performed at a temperature of -20 to 40 ° C. for 1 to 24 hours, preferably at a temperature of -10 to 30 ° C. for 1 to 12 hours, more preferably. Specifically, it may be performed while stirring at a temperature of 0 to 10°C for 1 to 6 hours. If the reaction temperature is less than -10°C, the reaction temperature is too low and stirring may be difficult, and if the reaction temperature exceeds 40°C, the cyclization reaction proceeds, resulting in an increase in impurities.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제3단계의 티올계 화합물은 알킬티올, 방향족 티올 또는 티오글리콜산을 의미할 수 있으며, 상기 알킬티올의 알킬기는 C1~C8의 직쇄형 알킬기를 의미할 수 있다.In the method for producing ectoine according to the present invention, the thiol-based compound in the third step may mean alkylthiol, aromatic thiol, or thioglycolic acid, and the alkyl group of the alkylthiol is a straight-chain alkyl group of C1 to C8. It can mean.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 알킬아세트이미데이트 화합물은 하기와 같은 화학식 3의 화합물을 의미할 수 있다.In the method for producing ectoine according to the present invention, the alkylacetimidate compound may refer to a compound of formula 3 below.
[화학식 3][Formula 3]
CH3C(=NH)OR · HClCH 3 C(=NH)OR · HCl
상기 화학식 3에서, R은 C1~C8의 직쇄형 알킬기를 의미할 수 있다.In Formula 3, R may refer to a straight-chain alkyl group of C1 to C8.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제4단계의 염기는 유기염기, 무기염기 또는 이들의 혼합물을 의미할 수 있으며, 상기 유기염기는 트리에틸아민, 다이이소프로필에틸아민, 다이메틸아민 및 다이에틸아민으로 이루어진 군으로부터 선택된 1종 이상을 의미할 수 있다.In the method for producing ectoine according to the present invention, the base in the fourth step may mean an organic base, an inorganic base, or a mixture thereof, and the organic base is triethylamine, diisopropylethylamine, dimethyl It may refer to one or more types selected from the group consisting of amine and diethylamine.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제4단계의 무기염기는 소듐 t-부톡사이드, 포타슘 t-부톡사이드, 소듐 하이드라이드, 소듐 메톡사이드, 소듐 에톡사이드, 리튬 하이드라이드, 리튬 하이드록사이드, 리튬 t-부톡사이드, 리튬 메톡사이드, 리튬 에톡사이드, 소듐 하이드록사이드 및 포타슘 하이드록사이드로 이루어진 군으로부터 선택된 1종 이상 또는 이들의 혼합물을 의미할 수 있다.In the method for producing ectoine according to the present invention, the inorganic base in the fourth step is sodium t-butoxide, potassium t-butoxide, sodium hydride, sodium methoxide, sodium ethoxide, lithium hydride, and lithium hydride. It may mean one or more selected from the group consisting of oxide, lithium t-butoxide, lithium methoxide, lithium ethoxide, sodium hydroxide, and potassium hydroxide, or a mixture thereof.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제4단계의 유기염기 또는 무기염기는 상기 (S)-2,4-디아미노 부타노익산 1 당량에 대하여 0.9 내지 3.0 당량비, 바람직하게는 1.0 내지 2.5 당량비, 더욱 바람직하게는 1.0 내지 2.0 당량비를 첨가하는 것을 특징으로 할 수 있다.In the method for producing ectoine according to the present invention, the organic base or inorganic base in the fourth step is used in an equivalent ratio of 0.9 to 3.0, preferably 1.0, based on 1 equivalent of (S)-2,4-diaminobutanoic acid. It may be characterized by adding an equivalent ratio of from 1.0 to 2.5, more preferably from 1.0 to 2.0.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제4단계의 알코올계 유기용매는 메탄올, 에탄올, 프로판올, 이소프로판올, n-부탄올, 이소부탄올, sec-부탄올, tert-부탄올, 펜탄올, 헥산올, 헵탄올, 옥탄올, 아이소옥탄올 및 2-에틸헥산올로 이루어진 군으로부터 선택된 1종 이상 또는 이들의 혼합물을 의미할 수 있다.In the method for producing ectoine according to the present invention, the alcohol-based organic solvent in the fourth step is methanol, ethanol, propanol, isopropanol, n-butanol, isobutanol, sec-butanol, tert-butanol, pentanol, and hexanol. , it may mean one or more types selected from the group consisting of heptanol, octanol, isooctanol, and 2-ethylhexanol, or a mixture thereof.
한편, 본 발명에 따른 엑토인의 제조방법에 있어서, 각 단계에서 생성되는 중간체들은 별도로 분리하여 염을 제조할 수 있다.Meanwhile, in the method for producing ectoine according to the present invention, the intermediates produced in each step can be separated separately to prepare a salt.
구체적으로, 본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제2단계에 의해 수득되는 상기 제2중간체에 염산, 브롬산, 술폰산, 벤젠술폰산, 켐퍼술폰산, 아세트산, 트리플루오로 아세트산, 메탄설폰산, 인산 및 유기산으로 이루어진 군으로부터 선택된 1종 이상 또는 이들의 혼합물에 해당하는 산을 투입하여 염((S)-4-아미노-(2-nonyl)부타노익산 염)을 제조할 수 있다.Specifically, in the method for producing ectoine according to the present invention, the second intermediate obtained in the second step includes hydrochloric acid, hydrobromic acid, sulfonic acid, benzenesulfonic acid, kaempfersulfonic acid, acetic acid, trifluoroacetic acid, and methane sulfur. A salt ((S)-4-amino-(2-nonyl)butanoic acid salt) can be prepared by adding one or more acids selected from the group consisting of phonic acid, phosphoric acid, and organic acid, or a mixture thereof.
구체적으로, 본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제3단계에 의해 수득되는 상기 (S)-2,4-디아미노부타노익산에 염산, 황산, 초산, 인산, 트리플루오로아세트산, 브롬산, 벤젠술폰산, 켐퍼술폰산, 및 유기산으로 이루어진 군으로부터 선택된 1종 이상 또는 이들의 혼합물에 해당하는 산을 투입하여 염((S)-2,4-디아미노부타노익산 염)을 제조할 수 있다.Specifically, in the method for producing ectoine according to the present invention, hydrochloric acid, sulfuric acid, acetic acid, phosphoric acid, and trifluoroacetic acid are added to the (S)-2,4-diaminobutanoic acid obtained in the third step. , prepare a salt ((S)-2,4-diaminobutanoic acid salt) by adding one or more acids selected from the group consisting of bromic acid, benzenesulfonic acid, kaempfersulfonic acid, and organic acids, or a mixture thereof. can do.
본 발명에 따른 엑토인의 제조방법은 상기 제4단계 이후 추가적으로 결정화 공정을 수행할 수 있으며, 이때 상기 결정화 공정의 경우 직쇄 및 분쇄형 C1~C8 알코올류, 메틸아세테이트, 에틸아세테이트, 아이소프로필아세테이트, 부틸아세테이트, 아세토니트릴, 테트라하이드로퓨란, 메틸렌클로라이드 및 아세톤으로 이루어진 군으로부터 선택된 1종 이상을 용매로 사용할 수 있다.The method for producing ectoine according to the present invention can additionally carry out a crystallization process after the fourth step. In this case, in the case of the crystallization process, straight chain and crushed C1 to C8 alcohols, methyl acetate, ethyl acetate, isopropyl acetate, One or more types selected from the group consisting of butylacetate, acetonitrile, tetrahydrofuran, methylene chloride, and acetone may be used as the solvent.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제4단계는 60 내지 180℃의 온도에서 1 내지 24시간 동안, 바람직하게는 60 내지 150℃의 온도에서 1 내지 18시간 동안, 더욱 바람직하게는 65 내지 120℃의 온도에서 1 내지 12시간 동안 교반시키면서 수행하는 것이 좋으며, 이때 반응온도가 130℃을 초과하는 경우 최종적으로 수득되는 물질의 색이 어두워지는 결과가 발생하는 문제점이 있다.In the method for producing ectoine according to the present invention, the fourth step is performed at a temperature of 60 to 180 ° C. for 1 to 24 hours, preferably at a temperature of 60 to 150 ° C. for 1 to 18 hours, more preferably It is recommended to carry out the reaction at a temperature of 65 to 120°C while stirring for 1 to 12 hours. However, if the reaction temperature exceeds 130°C, there is a problem in that the color of the final material obtained becomes dark.
본 발명에 따른 엑토인의 제조방법에 있어서, 최종적으로 수득되는 상기 엑토인은 유리염기 또는 금속염일 수 있으며, 상기 금속은 나트륨, 칼륨 또는 칼슘을 의미할 수 있으며, 이들의 수화물 또는 이들의 유기솔베이트를 포함할 수 있다.In the method for producing ectoine according to the present invention, the ectoine finally obtained may be a free base or a metal salt, and the metal may mean sodium, potassium, or calcium, and their hydrates or organic sols thereof. May include bait.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 금속염의 상기 금속은 알칼리금속, 알칼리토금속, 전이금속 또는 란탄족금속일 수 있다.In the method for producing ectoine according to the present invention, the metal of the metal salt may be an alkali metal, an alkaline earth metal, a transition metal, or a lanthanide metal.
본 발명에 따른 엑토인의 제조방법에 있어서, 상기 제4단계에서 생성된 엑토인에 염산, 황산, 초산, 인산, 트리플루오로아세트산, 브롬산, 벤젠술폰산, 켐퍼술폰산 및 유기산으로 이루어진 군으로부터 선택된 1종 이상 또는 이들의 혼합물에 해당하는 산을 투입하여 염을 제조할 수 있다.In the method for producing ectoine according to the present invention, the ectoine produced in the fourth step is mixed with an organic acid selected from the group consisting of hydrochloric acid, sulfuric acid, acetic acid, phosphoric acid, trifluoroacetic acid, hydrobromic acid, benzenesulfonic acid, kaempfersulfonic acid and organic acid. Salts can be prepared by adding one or more acids or a mixture thereof.
본 발명에 따른 엑토인의 제조방법은 하기 수학식 1에 의거 계산시, 전체수율이 45% 이상이며 순도가 99% 이상인 엑토인 화합물을 최종적으로 수득할 수 있다.The method for producing ectoine according to the present invention can ultimately obtain an ectoine compound with an overall yield of 45% or more and a purity of 99% or more when calculated according to Equation 1 below.
[수학식 1][Equation 1]
수율(%) = (반응생성물의 실제 수득량/반응생성물의 이론적 수득량) X 100%Yield (%) = (actual yield of reaction product/theoretical yield of reaction product)
또한, 위와 같은 공정을 수행하여 제조한 엑토인은 액체 크로마토그래피 분석 시 98% 이상, 바람직하게는 98 내지 99.9%, 더욱 바람직하게는 99 내지 99.9%의 높은 순도를 확보할 수 있다.In addition, ectoine prepared by performing the above process can secure a high purity of 98% or more, preferably 98 to 99.9%, and more preferably 99 to 99.9% when analyzed by liquid chromatography.
이하, 제조예들을 포함하는 구체적인 실시예들을 통해서 본 발명을 보다 상세하게 설명하기로 한다. 하지만, 이들은 본 발명을 보다 상세하게 설명하기 위한 것일 뿐 본 발명의 권리범위가 이에 한정되는 것이 아님은 자명할 것이다.Hereinafter, the present invention will be described in more detail through specific examples including manufacturing examples. However, it will be obvious that these are only for explaining the present invention in more detail and that the scope of the present invention is not limited thereto.
[실시예 1] 상기 화합물 2a로 표시되는 화합물인 (2-나이트로페닐)설포닐-L-글루타민의 제조[Example 1] Preparation of (2-nitrophenyl)sulfonyl-L-glutamine, a compound represented by compound 2a
1L의 이중자켓 반응조에 냉매를 순환시킨 후 시중에서 구입한 L-글루타민 38.3g, 산화마그네슘 32.6g, (2-나이트로벤젠설포닐)클로라이드 58.1g, 정제수 380mL를 차례대로 투입한 후 내부온도 25℃ 이하에서 2시간 동안 교반한다. 반응 종결을 TLC로 확인한 후 노란색 불투명한 반응액에 1N 수산화나트륨 용액을 가하여 pH를 10으로 조절한다. 녹지 않은 잔여 고체들을 셀라이트를 이용하여 여과하고 증류수 77mL로 세척한다. 노란색 여액에 염산을 가하여 pH를 1로 조절한 뒤 1시간 동안 교반한다. 생성된 고체를 여과하고 얼음물로 씻어준다. 열풍건조기에서 60℃에서 12시간 동안 건조하여 78.1의 화합물을 수득하였다(수율 90%). 이때, 수율은 상기 수학식 1에 의거하여 측정한 것이다. 1H NMR과 13C NMR의 측정결과는 하기와 같으며, 도 1 및 도 2에 도시하였다. 화합물 2b, 2c는 (2-나이트로벤젠설포닐)클로라이드 대신에 (4-나이트로벤젠설포닐)클로라이드, (2,4-다이나이트로벤젠설포닐)클로라이드를 사용하여 위와 동일한 방법으로 제조하였다. 1H NMR과 13C NMR의 측정결과는 하기와 같으며, 도 3 내지 도 6에 도시하였다.After circulating the refrigerant in a 1L double jacket reaction tank, 38.3g of commercially purchased L-glutamine, 32.6g of magnesium oxide, 58.1g of (2-nitrobenzenesulfonyl)chloride, and 380mL of purified water were sequentially added, and the internal temperature was 25. Stir for 2 hours at ℃ or lower. After confirming the completion of the reaction by TLC, 1N sodium hydroxide solution was added to the yellow, opaque reaction solution to adjust the pH to 10. The remaining undissolved solids were filtered using Celite and washed with 77 mL of distilled water. Add hydrochloric acid to the yellow filtrate to adjust the pH to 1 and stir for 1 hour. Filter the resulting solid and wash with ice water. By drying in a hot air dryer at 60°C for 12 hours, compound 78.1 was obtained (yield 90%). At this time, the yield was measured based on Equation 1 above. The measurement results of 1 H NMR and 13 C NMR are as follows and are shown in Figures 1 and 2. Compounds 2b and 2c were prepared in the same manner as above using (4-nitrobenzenesulfonyl)chloride and (2,4-dinitrobenzenesulfonyl)chloride instead of (2-nitrobenzenesulfonyl)chloride. . The measurement results of 1 H NMR and 13 C NMR are as follows and are shown in Figures 3 to 6.
<화합물 2a(흰색 고체)><Compound 2a (white solid)>
1H NMR(DMSO-d6) : δ 8.14-8.10(m, 1H), δ 8.04-7.97(m, 1H), δ 7.94-7.85(m, 2H), δ 3.41-3.39(t, 1H), δ 2.95-2.80(m, 2H), δ 2.05-1.85(m, 2H) 1 H NMR (DMSO-d6): δ 8.14-8.10 (m, 1H), δ 8.04-7.97 (m, 1H), δ 7.94-7.85 (m, 2H), δ 3.41-3.39 (t, 1H), δ 2.95-2.80(m, 2H), δ 2.05-1.85(m, 2H)
13C NMR(DMSO-d6) : δ 171.23, δ 148.09, δ 134.73, δ 133.42, δ 132.43, δ 130.72, δ 125.51, δ 56.85, δ 37.01, δ 32.18 13 C NMR (DMSO-d6): δ 171.23, δ 148.09, δ 134.73, δ 133.42, δ 132.43, δ 130.72, δ 125.51, δ 56.85, δ 37.01, δ 32.18
<화합물 2b(수율 87%, 흰색 고체)><Compound 2b (87% yield, white solid)>
1H NMR(DMSO-d6) : δ 8.61-8.59(d, 1H), δ 8.41-8.39(d, 2H), δ 8.03-8.01(d, 2H), δ 7.27(s, 1H), δ 6.77(s, 1H), δ 3.85-3.75(m, 1H), δ 3.10-2.07(t, 2H), δ 1.91-1.83(m, 1H), δ 1.7-1.6(m, 1H) 1 H NMR (DMSO-d6): δ 8.61-8.59 (d, 1H), δ 8.41-8.39 (d, 2H), δ 8.03-8.01 (d, 2H), δ 7.27 (s, 1H), δ 6.77 ( s, 1H), δ 3.85-3.75 (m, 1H), δ 3.10-2.07 (t, 2H), δ 1.91-1.83 (m, 1H), δ 1.7-1.6 (m, 1H)
13C NMR(DMSO-d6) : δ 173.45, δ 172.82, δ 149.88, δ 147.21, δ 128.56, δ 124.83, δ 55.83, δ 31.09, δ 28.12 13 C NMR (DMSO-d6): δ 173.45, δ 172.82, δ 149.88, δ 147.21, δ 128.56, δ 124.83, δ 55.83, δ 31.09, δ 28.12
<화합물 2c(수율 81%, 노란색 고체)><Compound 2c (81% yield, yellow solid)>
1H NMR(DMSO-d6) : δ 9.03-9.01(d, 1H), δ 8.90(s, 1H), δ 8.32-8.29(d, 1H), δ 7.37(s, 1H), δ 7.29-7.27(d, 1H), δ 6.83(s, 1H), δ 4.75-4.65(m, 1H), δ 2.21-2.02(m, 4H) 1 H NMR (DMSO-d6): δ 9.03-9.01 (d, 1H), δ 8.90 (s, 1H), δ 8.32-8.29 (d, 1H), δ 7.37 (s, 1H), δ 7.29-7.27 ( d, 1H), δ 6.83 (s, 1H), δ 4.75-4.65 (m, 1H), δ 2.21-2.02 (m, 4H)
13C NMR(DMSO-d6) : δ 173.78, δ 172.68, δ 147.56, δ 135.94, δ 130.78, δ 130.60, δ 124.00, δ 116.26, δ 55.23, δ 30.54, δ 27.09 13 C NMR (DMSO-d6): δ 173.78, δ 172.68, δ 147.56, δ 135.94, δ 130.78, δ 130.60, δ 124.00, δ 116.26, δ 55.23, δ 30.54, δ 27.09
[실시예 2] 상기 화합물 3a로 표시되는 화합물인 (S)-4-아미노-2-(2-나이트로페닐)부타노익산의 제조[Example 2] Preparation of (S)-4-amino-2-(2-nitrophenyl)butanoic acid, a compound represented by compound 3a.
1L의 이중자켓 반응조에 냉매를 순환시킨 후 (2-나이트로페닐설포닐)-L-글루타민 66.3g, 아세토나이트릴 200mL, 에틸아세테이트 100mL, 증류수 180mL를 차례대로 투입하고 내부온도 5℃ 이하로 냉각한다. 다음으로, (다이아세톡시아이오도)벤젠(PIDA) 70.9g을 3회로 소분하여 30분 동안 반응액에 투입하고 내부온도 0~5℃에서 5시간 동안 교반한다. 반응 종결을 TLC로 확인한 후 감압농축기로 반응액을 농축한 후 반응액을 1L 분액깔대기로 옮기고 에틸아세테이트 400mL를 넣고 혼합한 후 층분리를 수행한다. 수층을 40℃ 이하에서 감압농축한 후 잔류물에 메틸렌클로라이드 100mL를 넣고 상온에서 1시간동안 교반 후 여과하여 백색의 (S)-4-아미노-2-(2-나이트로페닐)부타노익산 48.5g을 수득하였다(수율 80%). 이때, 수율은 상기 수학식 1에 의거하여 측정한 것이다. 1H NMR과 13C NMR의 측정결과는 하기와 같으며, 도 7 및 도 8에 도시하였다. 화합물 3b, 3c에 관한 사항은 상기 실시예 1 및 실시예 2에 해당되는 사항과 동일한 방법으로 제조하였고, 1H NMR과 13C NMR의 측정결과는 하기와 같으며, 도 9 내지 도 12에 도시하였다.After circulating the refrigerant in a 1L double jacket reaction tank, 66.3g of (2-nitrophenylsulfonyl)-L-glutamine, 200mL of acetonitrile, 100mL of ethyl acetate, and 180mL of distilled water were sequentially added and cooled to an internal temperature of 5°C or lower. do. Next, 70.9 g of (diacetoxyiodo)benzene (PIDA) was divided into three portions and added to the reaction solution for 30 minutes, and stirred for 5 hours at an internal temperature of 0 to 5°C. After confirming the completion of the reaction by TLC, concentrate the reaction solution using a reduced pressure concentrator, transfer the reaction solution to a 1L separatory funnel, add 400 mL of ethyl acetate, mix, and perform layer separation. After concentrating the aqueous layer under reduced pressure below 40℃, 100 mL of methylene chloride was added to the residue, stirred at room temperature for 1 hour, and filtered to produce white (S)-4-amino-2-(2-nitrophenyl)butanoic acid 48.5. g was obtained (yield 80%). At this time, the yield was measured based on Equation 1 above. The measurement results of 1 H NMR and 13 C NMR are as follows and are shown in Figures 7 and 8. Compounds 3b and 3c were prepared in the same manner as in Examples 1 and 2 above, and the measurement results of 1 H NMR and 13 C NMR are as follows and are shown in Figures 9 to 12. did.
<화합물 3a(흰색 고체)><Compound 3a (white solid)>
1H NMR(DMSO-d6) : δ 8.14-8.10(m, 1H), δ 8.04-7.97(m, 1H), δ 7.94-7.85(m, 2H), δ 3.41-3.39(t, 1H), δ 2.95-2.80(m, 2H), δ 2.05-1.85(m, 2H) 1 H NMR (DMSO-d6): δ 8.14-8.10 (m, 1H), δ 8.04-7.97 (m, 1H), δ 7.94-7.85 (m, 2H), δ 3.41-3.39 (t, 1H), δ 2.95-2.80(m, 2H), δ 2.05-1.85(m, 2H)
13C NMR(DMSO-d6) : δ 171.23, δ 148.09, δ 134.73, δ 133.42, δ 132.43, δ 130.72, δ 125.51, δ 56.85, δ 37.01, δ 32.18 13 C NMR (DMSO-d6): δ 171.23, δ 148.09, δ 134.73, δ 133.42, δ 132.43, δ 130.72, δ 125.51, δ 56.85, δ 37.01, δ 32.18
<화합물 3b(수율 72%, 흰색 고체)><Compound 3b (yield 72%, white solid)>
1H NMR(DMSO-d6) : δ 8.37-8.35(d, 2H), δ 8.07-8.04(d, 2H), δ 3.27-3.24(t, 1H), δ 2.70-2.60(m, 2H), δ 1.77-1.61(m, 2H) 1 H NMR (DMSO-d6): δ 8.37-8.35 (d, 2H), δ 8.07-8.04 (d, 2H), δ 3.27-3.24 (t, 1H), δ 2.70-2.60 (m, 2H), δ 1.77-1.61(m, 2H)
13C NMR(DMSO-d6) : δ 172.00, δ 149.81, δ 146.72, δ 128.82, δ 124.71, δ 56.52, δ 38.54, δ 37.46 13 C NMR (DMSO-d6): δ 172.00, δ 149.81, δ 146.72, δ 128.82, δ 124.71, δ 56.52, δ 38.54, δ 37.46
<화합물 3c(수율 71%, 노란색 고체)><Compound 3c (71% yield, yellow solid)>
1H NMR(DMSO-d6) : δ 9.63(s, br, 1H), δ 8.88(s, 1H), δ 8.23-8.21(d, 1H), δ 7.10-7.08(d, 1H), δ 4.10-4.08(d, 1H), δ 3.15-3.08(m, 1H), δ 2.99-2.90(m, 1H), δ 2.10-1.99(m, 1H), δ 1.97-1.80(m, 1H) 1 H NMR (DMSO-d6): δ 9.63 (s, br, 1H), δ 8.88 (s, 1H), δ 8.23-8.21 (d, 1H), δ 7.10-7.08 (d, 1H), δ 4.10- 4.08 (d, 1H), δ 3.15-3.08 (m, 1H), δ 2.99-2.90 (m, 1H), δ 2.10-1.99 (m, 1H), δ 1.97-1.80 (m, 1H)
13C NMR(DMSO-d6) : δ 171.80, δ 146.65, δ 134.98, δ 130.41, δ 124.33, δ 116.50, δ 56.19, δ 37.27, δ 29.55 13 C NMR (DMSO-d6): δ 171.80, δ 146.65, δ 134.98, δ 130.41, δ 124.33, δ 116.50, δ 56.19, δ 37.27, δ 29.55
[실시예 3] (S)-2,4-디아미노부타노익산의 제조[Example 3] Preparation of (S)-2,4-diaminobutanoic acid
1000mL 2구 라운드 플라스크에 콘덴서를 연결하고 냉매를 순환시킨 후 상기 실시예 2에서 제조한 상기 화합물 3a 30.33g, 티오페놀 27.54g, 포타슘 하이드록사이드 14.03g, 아세토니트릴 150mL, 증류수 200mL를 투입하고 2시간 동안 55℃에서 교반하였다. 다음으로, 반응 종결을 TLC로 확인하고 반응액을 상온으로 냉각한 후 1M 염산용액으로 pH를 7로 조절하고 감압농축하여 용매를 제거한다. 농축잔류물에 아이소프로필에테르를 200mL를 넣고 상온에서 2시간 동안 교반 후 여과를 실시하여 (S)-2,4-디아미노부타노익산 10.75g을 수득하였다(수율 91%). 화합물 3b, 3c는 위와 동일한 방법으로 수행하여 각각 수율 92%, 94%로 수득하였다. 1H NMR과 13C NMR의 측정결과는 하기와 같으며, 도 13 및 도 14에 도시하였다. After connecting the condenser to a 1000 mL two-necked round flask and circulating the refrigerant, 30.33 g of the compound 3a prepared in Example 2, 27.54 g of thiophenol, 14.03 g of potassium hydroxide, 150 mL of acetonitrile, and 200 mL of distilled water were added. Stirred at 55°C for 1 hour. Next, the completion of the reaction is confirmed by TLC, the reaction solution is cooled to room temperature, the pH is adjusted to 7 with 1M hydrochloric acid solution, and the solvent is removed by concentration under reduced pressure. 200 mL of isopropyl ether was added to the concentrated residue, stirred at room temperature for 2 hours, and then filtered to obtain 10.75 g of (S)-2,4-diaminobutanoic acid (yield 91%). Compounds 3b and 3c were obtained in the same manner as above with yields of 92% and 94%, respectively. The measurement results of 1 H NMR and 13 C NMR are as follows and are shown in Figures 13 and 14.
1H NMR(D2O) : δ 3.75-3.71(t, 1H), δ 3.16-3.03(m, 2H), δ 2.13-2.08(q, 2H) 1 H NMR (DO): δ 3.75-3.71 (t, 1H), δ 3.16-3.03 (m, 2H), δ 2.13-2.08 (q, 2H)
13C NMR(D2O) : δ 172, δ 51.58, δ 36.55, δ 27.92 13 C NMR(DO): δ 172, δ 51.58, δ 36.55, δ 27.92
[실시예 4] 엑토인의 제조-1[Example 4] Preparation of ectoine-1
500mL 2구 라운드 플라스크에 콘덴서를 연결하고 냉매를 순환시킨 후 상기 (S)-2,4-디아미노부타노익산 11.81g, n-부탄올 120mL, 트리메틸오쏘아세테이트(trimethylorthoacetate) 24.1g을 투입하고 12시간 동안 환류교반하였다. 다음으로, 반응 종결을 TLC로 확인하고 반응액을 상온으로 냉각한 후 감압농축하여 용매를 제거한다. 잔류물에 메탄올 50mL와 에틸아세테이트 100mL를 넣고 상온에서 2시간 동안 교반한 후 여과를 실시하여 최종적으로 엑토인 10.24g을 획득하였다(수율 72%). 또한 수득된 엑토인을 액체 크로마토그래피로 분석하여 순도가 99.3%을 확인하였고, 1H NMR과 13C NMR의 측정결과는 하기와 같으며, 도 15 및 도 16에 도시하였다.Connect the condenser to a 500mL two-necked round flask, circulate the refrigerant, add 11.81g of (S)-2,4-diaminobutanoic acid, 120mL of n-butanol, and 24.1g of trimethylorthoacetate and leave for 12 hours. It was refluxed and stirred for a while. Next, completion of the reaction is confirmed by TLC, the reaction solution is cooled to room temperature, and the solvent is removed by concentration under reduced pressure. 50 mL of methanol and 100 mL of ethyl acetate were added to the residue, stirred at room temperature for 2 hours, and then filtered to obtain 10.24 g of ectoin (yield 72%). In addition, the obtained ectoine was analyzed by liquid chromatography to confirm that the purity was 99.3%, and the measurement results of 1 H NMR and 13 C NMR were as follows and are shown in Figures 15 and 16.
1H NMR(D2O)= δ 3.95-3.92(t, 1H), δ 3.35-3.30(m, 1H), δ 3.20-3.12(m, 1H), δ 2.10(s, 3H), δ 2.04-1.93(m, 2H) 1 H NMR(DO)=δ 3.95-3.92(t, 1H), δ 3.35-3.30(m, 1H), δ 3.20-3.12(m, 1H), δ 2.10(s, 3H), δ 2.04-1.93( m, 2H)
13C NMR(D2O)= δ 176.66, δ 160.40, δ 53.08, δ 37.11, δ 21.30, δ 18.06 13 C NMR(DO)=δ 176.66, δ 160.40, δ 53.08, δ 37.11, δ 21.30, δ 18.06
[실시예 5] 엑토인의 제조-2[Example 5] Preparation of ectoine-2
500mL 2구 라운드 플라스크에 콘덴서를 연결하고 냉매를 순환시킨 후 상기 (S)-2,4-디아미노부타노익산 11.81g, n-부탄올 120mL, 에틸아세트이미데이트 염산 13.6g, 트리에틸아민 20.24g을 투입하고 24시간 동안 환류교반하였다. 다음으로 반응 종결을 TLC로 확인하고 반응액을 상온으로 냉각한 후 감압농축하여 용매를 제거한다. 잔류물에 메탄올 50mL와 아세톤 110mL를 넣고 한시간 동안 환류한 다음 상온에서 1시간 동안 교반 후 여과를 실시하여 최종적으로 엑토인 9.67g을 획득하였다(수율 68%).Connect the condenser to a 500mL two-necked round flask, circulate the refrigerant, and add 11.81g of (S)-2,4-diaminobutanoic acid, 120mL of n-butanol, 13.6g of ethyl acetimidate hydrochloric acid, and 20.24g of triethylamine. was added and stirred under reflux for 24 hours. Next, the completion of the reaction is confirmed by TLC, the reaction solution is cooled to room temperature, and the solvent is removed by concentrating under reduced pressure. 50 mL of methanol and 110 mL of acetone were added to the residue, refluxed for 1 hour, stirred at room temperature for 1 hour, and then filtered to obtain 9.67 g of ectoin (yield 68%).
이상에서, 출원인은 본 발명의 바람직한 실시예들을 설명하였지만, 이와 같은 실시예들은 본 발명의 기술적 사상을 구현하는 일 실시예일 뿐이며, 본 발명의 기술적 사상을 구현하는 한 어떠한 변경례 또는 수정례도 본 발명의 범위에 속하는 것으로 해석되어야 할 것은 자명할 것이다.In the above, the applicant has described preferred embodiments of the present invention, but such embodiments are only one embodiment that implements the technical idea of the present invention, and no changes or modifications are permitted in this invention as long as the technical idea of the present invention is implemented. It will be obvious that it should be interpreted as falling within the scope of the invention.
Claims (5)
[화학식 1]
(상기 화학식 1에서, R1은 NO2 또는 H이고, R2는 NO2 또는 H이다)
(ⅱ) 상기 제1단계에서 수득한 상기 제1중간체 및 하이퍼발란트 아이오딘 시약(hypervalent iodine reagent)을 포함하는 혼합물을 반응시키는 과정을 포함하는 공정을 통해 하기 화학식 2로 표시되는 제2중간체를 수득하는 제2단계;
[화학식 2]
(상기 화학식 2에서, R1은 NO2 또는 H이고, R2는 NO2 또는 H이다)
(ⅲ) 상기 제2단계에서 수득한 상기 제2중간체 및 티올계 화합물을 포함하는 혼합물을 염기조건 하의 유기용매에 투입, 반응시키는 과정을 포함하는 공정을 통해 하기 화학식 3으로 표시되는 (S)-2,4-디아미노부타노익산을 수득하는 제3단계;
[화학식 3]
(ⅳ) 상기 제3단계에서 수득한 상기 (S)-2,4-디아미노부타노익산, 알킬아세트이미데이트 화합물 및 염기를 포함하는 혼합물을 유기용매에 투입, 반응시키는 과정을 포함하는 공정을 통해 하기 화학식 4로 표시되는 엑토인(Ectoine)을 수득하는 제4단계;를 포함하는, 엑토인의 제조방법.
[화학식 4]
(i) a first step of obtaining a first intermediate represented by the following formula (1) through a process comprising reacting a mixture containing L-glutamine and nitro-substituted benzenesulfonyl chloride under basic conditions;
[Formula 1]
(In Formula 1, R1 is NO 2 or H, and R2 is NO 2 or H)
(ii) producing a second intermediate represented by the following formula (2) through a process comprising reacting a mixture containing the first intermediate and a hypervalent iodine reagent obtained in the first step. The second step of obtaining;
[Formula 2]
(In Formula 2, R1 is NO 2 or H, and R2 is NO 2 or H)
(iii) (S)- represented by the following formula 3 through a process including adding the mixture containing the second intermediate and the thiol-based compound obtained in the second step to reacting in an organic solvent under basic conditions. The third step of obtaining 2,4-diaminobutanoic acid;
[Formula 3]
(iv) A process comprising adding the mixture containing (S)-2,4-diaminobutanoic acid, alkylacetimidate compound, and base obtained in the third step to an organic solvent and reacting it. A method for producing ectoine, comprising a fourth step of obtaining ectoine represented by the following formula (4).
[Formula 4]
하기 화학식 5로 표시되는 화합물인 것을 특징으로 하는, 엑토인의 제조방법.
[화학식 5]
The method of claim 1, wherein the hypervalent iodine reagent in the second step is
A method for producing ectoine, characterized in that it is a compound represented by the following formula (5).
[Formula 5]
상기 하이퍼발란트 아이오딘 시약이 상기 제1중간체 대비 1.0 내지 3.0 당량비로 포함되어 있는 혼합물인 것을 특징으로 하는, 엑토인의 제조방법.
The method of claim 1, wherein the mixture in the second step is
A method for producing ectoine, characterized in that it is a mixture containing the hypervalent iodine reagent in an equivalent ratio of 1.0 to 3.0 relative to the first intermediate.
알킬티올, 방향족티올 또는 티오글리콜산인 것을 특징으로 하며,
상기 알킬티올의 알킬기는 탄소수 1~8의 직쇄형 알킬기 중 어느 하나인 것을 특징으로 하는, 엑토인의 제조방법.
The method of claim 1, wherein the thiol-based compound of the third step is
Characterized by being an alkylthiol, aromatic thiol, or thioglycolic acid,
A method for producing ectoine, wherein the alkyl group of the alkylthiol is any one of a straight-chain alkyl group having 1 to 8 carbon atoms.
상기 염기가 상기 (S)-2,4-디아미노부타노익산 대비 1.0 내지 3.0 당량비로 포함되어 있는 혼합물인 것을 특징으로 하는, 엑토인의 제조방법.The method of claim 1, wherein the mixture in the fourth step is
A method for producing ectoine, characterized in that it is a mixture containing the base in an equivalent ratio of 1.0 to 3.0 compared to the (S)-2,4-diaminobutanoic acid.
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| (비특허문헌 1) E. A. Galinski et al., Eur. J. Biochem.m 149(1985) pages 135-139. |
| (특허문헌 1) JP 03031265 A (Takeda Chem Ind Ltd), 1989/06/26 |
| (특허문헌 2) US 0953988 A (Merck Patent GmbH), 1993/12/14 |
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