CN103992389A - Method for solid cyclizing synthesis of desmopressin - Google Patents
Method for solid cyclizing synthesis of desmopressin Download PDFInfo
- Publication number
- CN103992389A CN103992389A CN201310412013.5A CN201310412013A CN103992389A CN 103992389 A CN103992389 A CN 103992389A CN 201310412013 A CN201310412013 A CN 201310412013A CN 103992389 A CN103992389 A CN 103992389A
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- trt
- desmopressin
- resin
- fmoc
- reaction
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- Granted
Links
- 108010000437 Deamino Arginine Vasopressin Proteins 0.000 title claims abstract description 56
- 229960004281 desmopressin Drugs 0.000 title claims abstract description 48
- NFLWUMRGJYTJIN-NXBWRCJVSA-N desmopressin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@@H](CSSCCC(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(N)=O)=O)CCC(=O)N)C1=CC=CC=C1 NFLWUMRGJYTJIN-NXBWRCJVSA-N 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 42
- 239000007787 solid Substances 0.000 title claims abstract description 9
- 230000015572 biosynthetic process Effects 0.000 title abstract description 6
- 238000003786 synthesis reaction Methods 0.000 title abstract description 6
- 239000011347 resin Substances 0.000 claims abstract description 56
- 229920005989 resin Polymers 0.000 claims abstract description 56
- 239000007788 liquid Substances 0.000 claims abstract description 23
- 238000005336 cracking Methods 0.000 claims abstract description 15
- 230000003647 oxidation Effects 0.000 claims abstract description 10
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 10
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 9
- 239000007791 liquid phase Substances 0.000 claims abstract description 8
- 125000006239 protecting group Chemical group 0.000 claims abstract description 5
- 102000001189 Cyclic Peptides Human genes 0.000 claims abstract description 4
- 108010069514 Cyclic Peptides Proteins 0.000 claims abstract description 4
- 239000006227 byproduct Substances 0.000 claims abstract description 4
- 239000002699 waste material Substances 0.000 claims abstract description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 52
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 41
- DTQVDTLACAAQTR-UHFFFAOYSA-N trifluoroacetic acid Substances OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 36
- 238000006243 chemical reaction Methods 0.000 claims description 31
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 22
- 239000012071 phase Substances 0.000 claims description 18
- 229940078916 carbamide peroxide Drugs 0.000 claims description 13
- AQLJVWUFPCUVLO-UHFFFAOYSA-N urea hydrogen peroxide Chemical compound OO.NC(N)=O AQLJVWUFPCUVLO-UHFFFAOYSA-N 0.000 claims description 13
- 238000007363 ring formation reaction Methods 0.000 claims description 10
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 9
- 230000035484 reaction time Effects 0.000 claims description 7
- 150000001875 compounds Chemical class 0.000 claims description 6
- 238000010532 solid phase synthesis reaction Methods 0.000 claims description 6
- HNKJADCVZUBCPG-UHFFFAOYSA-N thioanisole Chemical compound CSC1=CC=CC=C1 HNKJADCVZUBCPG-UHFFFAOYSA-N 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 238000010511 deprotection reaction Methods 0.000 claims description 4
- 230000001590 oxidative effect Effects 0.000 claims description 3
- 230000000903 blocking effect Effects 0.000 claims description 2
- 238000011084 recovery Methods 0.000 claims 1
- 230000009466 transformation Effects 0.000 claims 1
- 150000001413 amino acids Chemical class 0.000 abstract description 10
- 238000010168 coupling process Methods 0.000 abstract description 9
- 230000008878 coupling Effects 0.000 abstract description 8
- 238000005859 coupling reaction Methods 0.000 abstract description 8
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 abstract description 6
- 229910052740 iodine Inorganic materials 0.000 abstract description 6
- 239000011630 iodine Substances 0.000 abstract description 6
- 239000007800 oxidant agent Substances 0.000 abstract description 5
- NDKDFTQNXLHCGO-UHFFFAOYSA-N 2-(9h-fluoren-9-ylmethoxycarbonylamino)acetic acid Chemical compound C1=CC=C2C(COC(=O)NCC(=O)O)C3=CC=CC=C3C2=C1 NDKDFTQNXLHCGO-UHFFFAOYSA-N 0.000 abstract description 4
- KLBPUVPNPAJWHZ-UMSFTDKQSA-N (2r)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-tritylsulfanylpropanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)SC(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 KLBPUVPNPAJWHZ-UMSFTDKQSA-N 0.000 abstract description 2
- HNICLNKVURBTKV-MUUNZHRXSA-N (2r)-5-[[amino-[(2,2,4,6,7-pentamethyl-3h-1-benzofuran-5-yl)sulfonylamino]methylidene]amino]-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)N[C@@H](C(O)=O)CCCN=C(N)NS(=O)(=O)C1=C(C)C(C)=C2OC(C)(C)CC2=C1C HNICLNKVURBTKV-MUUNZHRXSA-N 0.000 abstract description 2
- ZPGDWQNBZYOZTI-SFHVURJKSA-N (2s)-1-(9h-fluoren-9-ylmethoxycarbonyl)pyrrolidine-2-carboxylic acid Chemical compound OC(=O)[C@@H]1CCCN1C(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 ZPGDWQNBZYOZTI-SFHVURJKSA-N 0.000 abstract description 2
- JAUKCFULLJFBFN-VWLOTQADSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[4-[(2-methylpropan-2-yl)oxy]phenyl]propanoic acid Chemical compound C1=CC(OC(C)(C)C)=CC=C1C[C@@H](C(O)=O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 JAUKCFULLJFBFN-VWLOTQADSA-N 0.000 abstract description 2
- SJVFAHZPLIXNDH-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-phenylpropanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C1=CC=CC=C1 SJVFAHZPLIXNDH-QFIPXVFZSA-N 0.000 abstract description 2
- KJYAFJQCGPUXJY-UMSFTDKQSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-oxo-4-(tritylamino)butanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C(=O)NC(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 KJYAFJQCGPUXJY-UMSFTDKQSA-N 0.000 abstract description 2
- WDGICUODAOGOMO-DHUJRADRSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-5-oxo-5-(tritylamino)pentanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)CC(=O)NC(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 WDGICUODAOGOMO-DHUJRADRSA-N 0.000 abstract description 2
- AECGEIVNZGQBJT-UHFFFAOYSA-N 3-tritylsulfanylpropanoic acid Chemical compound C=1C=CC=CC=1C(C=1C=CC=CC=1)(SCCC(=O)O)C1=CC=CC=C1 AECGEIVNZGQBJT-UHFFFAOYSA-N 0.000 abstract description 2
- 238000011031 large-scale manufacturing process Methods 0.000 abstract description 2
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 43
- 238000000967 suction filtration Methods 0.000 description 20
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 19
- 238000003756 stirring Methods 0.000 description 18
- 238000003746 solid phase reaction Methods 0.000 description 16
- 238000010671 solid-state reaction Methods 0.000 description 16
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 12
- 238000005406 washing Methods 0.000 description 12
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 150000001408 amides Chemical class 0.000 description 9
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 8
- 239000013558 reference substance Substances 0.000 description 8
- RDRBIXSNGAYLPT-UHFFFAOYSA-N CC1=CC=C(COC2=CC3=C(C=C2)C(NC(=O)OCC2C4=C(C=CC=C4)C4=C2C=CC=C4)C2=C(O3)C=CC=C2)C=C1 Chemical compound CC1=CC=C(COC2=CC3=C(C=C2)C(NC(=O)OCC2C4=C(C=CC=C4)C4=C2C=CC=C4)C2=C(O3)C=CC=C2)C=C1 RDRBIXSNGAYLPT-UHFFFAOYSA-N 0.000 description 7
- 229940024606 amino acid Drugs 0.000 description 7
- 235000001014 amino acid Nutrition 0.000 description 7
- 238000004128 high performance liquid chromatography Methods 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 239000007790 solid phase Substances 0.000 description 6
- FIEYHAAMDAPVCH-UHFFFAOYSA-N 2-methyl-1h-quinazolin-4-one Chemical compound C1=CC=C2NC(C)=NC(=O)C2=C1 FIEYHAAMDAPVCH-UHFFFAOYSA-N 0.000 description 5
- TZCYLJGNWDVJRA-UHFFFAOYSA-N 6-chloro-1-hydroxybenzotriazole Chemical compound C1=C(Cl)C=C2N(O)N=NC2=C1 TZCYLJGNWDVJRA-UHFFFAOYSA-N 0.000 description 5
- 229960002845 desmopressin acetate Drugs 0.000 description 5
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 5
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 238000004090 dissolution Methods 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- VXGGBPQPMISJCA-STQMWFEESA-N (2s)-2-[[(2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)propanoyl]amino]propanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O)C3=CC=CC=C3C2=C1 VXGGBPQPMISJCA-STQMWFEESA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 3
- 229910019142 PO4 Inorganic materials 0.000 description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 3
- 238000013016 damping Methods 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 230000002101 lytic effect Effects 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 3
- 239000010452 phosphate Substances 0.000 description 3
- 229910052700 potassium Inorganic materials 0.000 description 3
- 239000011591 potassium Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- REFMEZARFCPESH-UHFFFAOYSA-M sodium;heptane-1-sulfonate Chemical compound [Na+].CCCCCCCS([O-])(=O)=O REFMEZARFCPESH-UHFFFAOYSA-M 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 2
- 208000032843 Hemorrhage Diseases 0.000 description 2
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- BZLVMXJERCGZMT-UHFFFAOYSA-N Methyl tert-butyl ether Chemical compound COC(C)(C)C BZLVMXJERCGZMT-UHFFFAOYSA-N 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 125000005519 fluorenylmethyloxycarbonyl group Chemical group 0.000 description 2
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- CMWYAOXYQATXSI-UHFFFAOYSA-N n,n-dimethylformamide;piperidine Chemical compound CN(C)C=O.C1CCNCC1 CMWYAOXYQATXSI-UHFFFAOYSA-N 0.000 description 2
- QWVGKYWNOKOFNN-UHFFFAOYSA-N o-cresol Chemical compound CC1=CC=CC=C1O QWVGKYWNOKOFNN-UHFFFAOYSA-N 0.000 description 2
- 235000011837 pasties Nutrition 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- 238000004062 sedimentation Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- 230000002194 synthesizing effect Effects 0.000 description 2
- 238000010189 synthetic method Methods 0.000 description 2
- BDNKZNFMNDZQMI-UHFFFAOYSA-N 1,3-diisopropylcarbodiimide Chemical compound CC(C)N=C=NC(C)C BDNKZNFMNDZQMI-UHFFFAOYSA-N 0.000 description 1
- DKIDEFUBRARXTE-UHFFFAOYSA-N 3-mercaptopropanoic acid Chemical compound OC(=O)CCS DKIDEFUBRARXTE-UHFFFAOYSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical class N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- ODKSFYDXXFIFQN-SCSAIBSYSA-N D-arginine Chemical compound OC(=O)[C@H](N)CCCNC(N)=N ODKSFYDXXFIFQN-SCSAIBSYSA-N 0.000 description 1
- 206010051055 Deep vein thrombosis Diseases 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 208000031220 Hemophilia Diseases 0.000 description 1
- 208000009292 Hemophilia A Diseases 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 description 1
- 102100026383 Vasopressin-neurophysin 2-copeptin Human genes 0.000 description 1
- 206010047249 Venous thrombosis Diseases 0.000 description 1
- 125000004202 aminomethyl group Chemical group [H]N([H])C([H])([H])* 0.000 description 1
- 238000004176 ammonification Methods 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 239000012964 benzotriazole Substances 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 201000010064 diabetes insipidus Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 230000000025 haemostatic effect Effects 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 238000013094 purity test Methods 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 125000002221 trityl group Chemical group [H]C1=C([H])C([H])=C([H])C([H])=C1C([*])(C1=C(C(=C(C(=C1[H])[H])[H])[H])[H])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
The invention relates to a method for solid cyclizing synthesis of desmopressin. The method solves the technical problem that the existing air liquid-phase oxidation method produces a large amount of waste liquid, utilizes behindhand processes, has low efficiency and produces more by-products because of oxidation adopting iodine as an oxidizing agent. The method comprises the following steps of orderly coupling nine amino acids to a RinkAmideAm resin, wherein the coupled amino acids orderly comprise Fmoc-Gly-OH, Fmoc-D-Arg(pbf)-OH, Fmoc-Pro-OH, Fmoc-Cys(Trt)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Phe-OH, Fmoc-Tyr(tBu)-OH and Mpa(Trt)-OH, removing protective groups Trt on Cys(Trt) and Mpa(Trt) by TFA/DCM having content of 1%, carrying out solid cyclizing synthesis by an oxidation reagent to obtain a cyclopeptide resin and carrying out cracking to obtain desmopressin. The invention provides the method for solid cyclizing synthesis of desmopressin and the method has simple processes, a low cost and a high yield and is suitable for large-scale production.
Description
Technical field
The present invention relates to a kind of preparation method of polypeptide drugs, particularly a kind of solid phase synthesis process of Desmopressin.
Background technology
Desmopressin, English name is: Desmopressin is the analog of natural smart ammonia salt vassopressin, is that the chemical structure of natural hormone is carried out the change of two places and obtained, i.e. Cys deaminize and replace 8-L-arginine with 8-D-arginine.Desmopressin acetate has good haemostatic effect and can not produce the feature of deep venous thrombosis.Before being mainly used to treat hemophilia, diabetes insipidus and therapeutic Bleeding control and operation, prevention is hemorrhage.Effective and side effect is little, be a kind of polypeptide drugs of good market prospect.Its peptide sequence structure is as follows:
Mpa-Tyr-Phe-Gln-Asn-Cys-Pro-D-Arg-Gly-NH
2(Sulfide?bond?between?Mpa?and?Cys)
WO2011011342 discloses a kind of method of acetic acid synthesized Desmopressin on MBHA, Sieber and CTC resin respectively, the method adopts iodine/methylene dichloride to carry out solid phase cyclization, wherein CTC resin solid phase cyclization, cracking obtain the Desmopressin precursor that carbon teminal is carboxyl, carry out ammonification again and obtain desmopressin acetate, the purity of synthesizing products obtained therefrom by three kinds of resins and different methods is all lower than 90%.US5726287 discloses a kind of method that adopts respectively Boc method (mbha resin) and the acetic acid synthesized Desmopressin of Fmoc method (AM resin), concrete steps are: first acetic acid synthesized Desmopressin linear peptides, then under aqueous phase system and iodine oxidizing condition, carry out cyclisation and generate disulfide linkage, finally adopt respectively ion-exchange packing and reversed-phase HPLC to carry out purifying, product purity is all lower than 95%.CN101372505 discloses a kind of method of the Sieber of employing Amide resin solid phase synthesis desmopressin acetate, it passes through progressively coupling method and synthesizes after Desmopressin linear peptides resin, in acetonitrile/water system, with hydrogen peroxide, linear peptides is oxidized and generates Desmopressin peptide resin, through cracking, purifying, obtain desmopressin acetate again, its yield is 55%, and product purity has no report.But Sieber Amide resin is a kind of very special resin, supplier seldom, be difficult for buying, and price is high.It is synthetic as the Fmoc method that initial resin has carried out Desmopressin that patent CN102863513 be take Rink Amide AM resin, after synthesizing linear peptide peptide resin cracking, then generates disulfide linkage, do not report for work yield and product purity by the liquid phase hydrogen peroxide oxidation of linear peptides.Patent CN103102395 adopts the synthetic Desmopressin linear peptides of Sieber Amide resin, cracking obtains the thick peptide of linear peptides and after purifying, in water, carry out again oxidizing reaction and generate Desmopressin, finally by turning salt, obtain desmopressin acetate, purity is more than 99.5%, and yield can reach 52%.Its raw material Sieber Amide resin price costliness and the few shortcoming of supplier in depositing equally.
Summary of the invention
The object of this invention is to provide the processing method that a kind of solid cyclization becomes Desmopressin.The technical issues that need to address of the present invention are: (1) selects a kind of synthetic method of admittedly encircling, solve a large amount of waste liquids that liquid-phase oxidation brings, the problem that solid phase synthesis yield is low, (2) select a kind of oxygenant to substitute iodine, solve iodine many as the by product of oxygenant oxidation, the problem that yield is not high.
Preparation method of the present invention as shown in Figure 1, first 9 amino acid shown in Fig. 1 in coupling successively on AM resin, coupling amino acid is sequentially: the amino-acid sequence of coupling is: Fmoc-Gly-OH, Fmoc-D-Arg (pbf)-OH, Fmoc-Pro-OH, Fmoc-Cys (Trt)-OH, Fmoc-Asn (Trt)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Phe-OH, Fmoc-Tyr (tBu)-OH, Mpa (Trt)-OH, then use containing the DCM solution removal Cys (Trt) of 1%TFA and the protecting group Trt on Mpa (Trt), then adding oxidising agent admittedly encircles and obtains cyclic peptide resin, finally by cracking, obtain Desmopressin.
In the present invention, some conventional abbreviations have following implication;
Fmoc: fluorenylmethyloxycarbonyl
Fmoc-AA-OH: the amino acid of fluorenylmethyloxycarbonyl protection
TBu: the tertiary butyl
Trt: trityl
Pbf:2,2,4,6,7-pentamethyl-Dihydrobenzofuranes-5-alkylsulfonyl
The chloro-1-hydroxyl of Cl-HOBt: 6-benzotriazole
DIC: N, N '-di-isopropyl carbodiimide
Gly: glycine
D-Arg:D-type arginine
Pro: proline(Pro)
Cys: halfcystine
Asn: asparagine
Gln: glutamine
Phe: phenylalanine
Tyr: tyrosine
Mpa: 3-thiohydracrylic acid
DMF: N, N '-dimethyl formamide
Piperidine: hexahydropyridine
Fmoc-Rink Amide Am resin: with the Rink aminomethyl resin of Fmoc protection
TFA: trifluoracetic acid
PhSMe: thioanisole
PhOMe: methyl-phenoxide
MeOH: methyl alcohol
DCM: methylene dichloride
Kaiser detection reagent is: 5% ethanol solution of ninhydrin; Pyridine; 80% phenol ethanolic soln
Oxygenant is: carbamide peroxide (solvent: DMSO)
For this reason, the invention provides a kind of method that solid cyclization becomes Desmopressin, described method steps is as follows:
Step 1, the linear Desmopressin resin of solid phase synthesis;
Step 2, by Cys (Trt), Mpa(Trt) on protecting group Trt deprotection;
Step 3, admittedly encircles and obtains cyclic peptide resin with oxygenant;
Step 4, resin is removed in cracking and blocking group obtains Desmopressin.
Wherein, linear Desmopressin resin described in step 1, structure is as follows:
Mpa(Trt)-Tyr(tBu)-Phe-Gln(Trt)-Asn(Trt)-Cys(Trt)-Pro-D-Arg(pbf)-Gly-NH-Resin
Its synthetic method can adopt the solid phase synthesis technique of existing routine.
Wherein, deprotection is to use the synthetic gained linearity of reagent A and step 1 Desmopressin resin to mix to react described in step 2, and the reaction times is 15min, and temperature of reaction is 25 ℃, reaction repeated 4 times.Wherein the compound method of reagent A is as follows: 1mLTFA is joined in 100mL DCM.Wherein, TFA is trifluoroacetic acid, and DCM is methylene dichloride.
Wherein, the oxygenant B that oxidation is used described in step 3 is selected from: carbamide peroxide solution, and its usage quantity is 100mL, and the reaction times is 2 hours, and temperature of reaction is 25 ℃.The compound method of oxygenant B is: the DMSO(dimethyl sulfoxide (DMSO) that the carbamide peroxide of 3.80g is joined to 100mL) in solution, dissolve.
Wherein, the cracking agent C that cracking is used described in step 4 is selected from: thioanisole and trifluoroacetic acid, and its usage quantity is 350mL, and the reaction times is 2 hours, and temperature of reaction is 25 ℃.The compound method of cracking agent C is: by the PhSMe(thioanisole of 35mL) add in the trifluoroacetic acid of 665mL.
The present invention passes through the specific deprotecting regent of screening by use, oxidising agent, and lytic reagent, finds a kind of method of high yield, obtains a kind of highly purified Desmopressin.
Below data declaration beneficial effect of the present invention by experiment:
The screening of deprotecting regent (volume ratio):
TFA:DCM?=0.5:100；TFA:DCM?=1:100；TFA:DCM?=2:100；TFA:DCM?=4:100
Use above-mentioned condition to prepare object peptide, obtain respectively the Desmopressin of different yields and purity, result data is as follows:
| Deprotecting regent (volume ratio) | Yield | Purity |
| TFA:DCM =0.5:100 | 73% | 90.66% |
| TFA:DCM =1:100 | 80% | 94.22% |
| TFA:DCM =2:100 | 70% | 91.48% |
| TFA:DCM =4:100 | 68% | 88.32% |
The screening of oxidising agent:
Carbamide peroxide; Iodine
Use above-mentioned condition to prepare object peptide, obtain respectively the Desmopressin of different yields and purity, result data is as follows:
| Oxidising agent | Yield | Purity |
| Carbamide peroxide | 80% | 94.22% |
| Iodine | 75% | 79.21% |
The screening of lytic reagent (volume ratio):
PhSMe:TFA=5:95; PhOMe:TFA=5:95; Wherein PhSMe is thioanisole, and PhOMe is methyl-phenoxide.
Use above-mentioned condition to prepare object peptide, obtain respectively the Desmopressin of different yields and purity, result data is as follows:
| Lytic reagent | Yield | Purity |
| PhSMe:TFA=5:95 | 80% | 94.22% |
| PhOMe:TFA=5:95 | 72% | 87.51% |
Compared to the prior art method of the present invention has obvious advantage, and relevant contrast experiment is as follows:
One, yield comparison:
By the method for the inventive method and prior art, carry out yield calculating, result is as follows:
| Patent | Resinous type | Cyclisation mode | Yield |
| The technology of the present invention | Rink Amide AM resin | Solid phase cyclization | 65.1% |
| CN101372505 | Sieber Amide resin | Solid phase cyclization | 55 % |
| CN103102395 | Sieber Amide resin | Liquid phase cyclisation | 52 % |
| CN102863513 | Rink Amide AM resin | Liquid phase cyclisation | Not quite clear |
Two, purity comparison:
By the method for the inventive method and prior art, carry out purity testing, result is as follows:
| Patent | Resinous type | Cyclisation mode | Purity % |
| The technology of the present invention | Rink Amide AM resin | Solid phase cyclization | 97.43% |
| CN101372505 | Sieber Amide resin | Solid phase cyclization | Not quite clear |
| CN103102395 | Sieber Amide resin | Liquid phase cyclisation | 92.5% |
| CN102863513 | Rink Amide AM resin | Liquid phase cyclisation | Not quite clear |
Method for detecting purity is wherein as follows:
Chromatographic column: use Inertsil ODS-2, (5 μ m) (250 * 4.6mm) or quite post;
Mobile phase A is acetonitrile-methyl alcohol-solution I (15:10:75), and Mobile phase B is acetonitrile-methyl alcohol (60:40); Flow velocity is 1.2mL/min; Detection wavelength is 220nm; 60 ℃ of column temperatures.The original state of gradient is 100% mobile phase A, keeps 15 minutes, then in 10 minutes, Mobile phase B is increased to 18%, then in 5 minutes, Mobile phase B is increased to 38%, keeps 5 minutes.
The invention has the beneficial effects as follows: use solid ring, operation is simple, cost is low and yield is higher, is applicable to large-scale production; The less side products of using carbamide peroxide to be oxidized as oxygenant than iodine, purity and yield are higher.
Accompanying drawing explanation
Fig. 1 is synthetic route chart of the present invention;
Fig. 2 is the HPLC color atlas (chromatographic column: Kromasil 100-5 C18 (250mm * 4.6mm) chromatographic column or quite post of the Desmopressin that obtains of embodiment 2; Mobile phase A: damping fluid (3.4g potassium primary phosphate and 2.0g sodium heptanesulfonate are dissolved in 1L water and adjust pH to 4.50 ± 0.05 with phosphoric acid or sodium hydroxide); Mobile phase B: acetonitrile; Elution flow rate: 1ml/min; Detect wavelength: 220nm; Gradient:
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 78 | 22 |
| 20 | 78 | 22 |
| 35 | 66 | 34 |
| 35.1 | 78 | 22 |
| 40 | 78 | 22 |
Fig. 3 is the HPLC color atlas (chromatographic column: Kromasil 100-5 C18 (250mm * 4.6mm) chromatographic column or quite post of Desmopressin reference substance; Mobile phase A: damping fluid (3.4g potassium primary phosphate and 2.0g sodium heptanesulfonate are dissolved in 1L water and adjust pH to 4.50 ± 0.05 with phosphoric acid or sodium hydroxide); Mobile phase B: acetonitrile; Elution flow rate: 1ml/min; Detect wavelength: 220nm;
Gradient:
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 78 | 22 |
| 20 | 78 | 22 |
| 35 | 66 | 34 |
| 35.1 | 78 | 22 |
| 40 | 78 | 22 |
Fig. 4 is the HPLC color atlas (chromatographic column: Kromasil 100-5 C18 (250mm * 4.6mm) chromatographic column or quite post of the Desmopressin that obtains of embodiment 3; Mobile phase A: damping fluid (3.4g potassium primary phosphate and 2.0g sodium heptanesulfonate are dissolved in 1L water and adjust pH to 4.50 ± 0.05 with phosphoric acid or sodium hydroxide); Mobile phase B: acetonitrile; Elution flow rate: 1ml/min; Detect wavelength: 220nm; Gradient:
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 78 | 22 |
| 20 | 78 | 22 |
| 35 | 66 | 34 |
| 35.1 | 78 | 22 |
| 40 | 78 | 22 |
Embodiment
Further illustrate by the following examples the present invention.
Embodiment 1:
Oxygenant preparation: the carbamide peroxide of 56.56g is added in the DMSO of 1500 mL and dissolve.
Embodiment 2:
Step 1 Fmoc-Gly-RinkAmide-AM-Resin's is synthetic
Take Rink Amide AM resin (substitution degree 0.73mmol/g) (164.85 g, 1.0 equiv) and add in solid state reaction still, add DCM(1000mL), temperature control 10-30 ℃ is stirred swelling 30min, and suction filtration is removed liquid.Add again DMF(1000mL * 2) washing.Measure 20%(v/v, lower same) Piperidine/DMF solution (850 mL), add in solid state reaction still, temperature control 20-30 ℃ of stirring reaction 5min, suction filtration is removed liquid.Again measure 20%Piperidine/DMF solution (850 mL), add in solid state reaction still, temperature control 20-30 ℃ of stirring reaction 10min, suction filtration is removed liquid.Measure DMF(1000 mL * 6) add the rear suction filtration of solid state reaction still washing to remove liquid.Separately take Fmoc-Gly-OH(107.13 g, 3.0 equiv), Cl-HOBt(67.15 g, 3.3 equiv) add beaker.Measure DMF(450 mL) add beaker, stirring is bathed and is cooled to 10-15 ℃ with cryosel after dissolving completely, adds DIC(55.8 mL, 3.0 equiv), stir temperature control 10-25 ℃ of reaction 5min, then this reaction solution is joined in solid state reaction still, after 2h, suction filtration is removed liquid.Measure DMF(1000 mL * 3) add the rear suction filtration of solid state reaction still washing to remove liquid.Triketohydrindene hydrate detects and is negative.
Step 2
Mpa (Trt)-Tyr (tBu)-Phe-Gln (Trt)-Asn (Trt)-Cys (Trt)-Pro-D-Arg (pbf)-Gly-RinkAmide-AM-Resin's is synthetic
Step 1 product amino-acid resin is sloughed to Fmoc by the same way.Take Fmoc-AA-OH(3.0 equiv), Cl-HOBt(3.3 equiv) add beaker.Measure DMF/DCM=1/1 solvent (500 mL) and add beaker, stirring is bathed and is cooled to 5-10 ℃ with cryosel after dissolving completely, adds DIC(3.0 equiv), stir, temperature control 5-25 ℃ of reaction 5min, then joins this reaction solution in solid state reaction still, and after 2h, suction filtration is removed liquid.Measure DMF(1200 mL * 3) add the rear suction filtration of solid state reaction still washing to remove liquid.Triketohydrindene hydrate detects, as positive in result, repeats coupling once.After all amino acid couplings finish, with DMF(1200 mL * 6) wash 6 times.
Step 3
Mpa-Tyr (tBu)-Phe-Gln (Trt)-Asn (Trt)-Cys-Pro-D-Arg (pbf)-Gly-RinkAmide-AM-Resin's is synthetic
In step 2 gained peptide resin, add the DCM solution (1500mL) containing 1%TFA, stirring reaction 15min, suction filtration is removed liquid.Repeat again 3 times, carry out altogether going for 4 times protection operation.Use again DMF(1200 mL * 6) wash 6 times.
Step 4
Mpa-Tyr (tBu)-Phe-Gln (Trt)-Asn (Trt)-Cys-Pro-D-Arg (pbf)-Gly-RinkAmide-AM-Resin's (Sulfide bond between Mpa and Cys) is synthetic
Under lucifuge, in step 3 gained peptide resin, add the DMSO solution (1500mL) that contains carbamide peroxide (56.56g, 5eq), control 20-30 ℃ of reaction 30min of temperature.Suction filtration is removed liquid, then use DMF(1500mL successively * 3), MeOH(1200mL * 2), DCM(2000mL * 2), MeOH(1200mL * 2) washing resin.Take out Desmopressin peptide resin, after being dried, obtain 495.5g.
Step 5
Mpa-Tyr-Phe-Gln-Asn-Cys-Pro-D-Arg-Gly-NH
2(Sulfide bond between Mpa and Cys's) is synthetic
Measure TFA(4750 mL), PhSMe(250 mL) join successively in 10L reaction flask, stirring and evenly mixing, controls lysate temperature at 15-20 ℃.Step 4 gained peptide resin is all added to reaction flask.Control temperature of reaction 25-30 ℃, reaction 2h.Filter, and with TFA(300mL * 2) wash.Merging filtrate, puts to revolve and steams 25% left and right that is concentrated to original volume under evaporimeter.Gained concentrated solution is joined to sedimentation in the methyl tertiary butyl ether of-10～-5 ℃.Centrifugal, washing after 6 times the thick peptide of pasty state, through drying under reduced pressure, obtain thick peptide 130.8g.The thick peptide of gained is dissolved with pure water 4L, after fully stirring, filter, obtain thick peptide solution.Thick peptide solution is carried out to HPLC quantitative analysis, obtain the about 83.5g of object peptide, thick peptide HPLC purity 97.43%, synthesis yield 65.1%.
Embodiment 3:
Step 1 Fmoc-Gly-RinkAmide-AM-Resin's is synthetic
Take Rink Amide AM resin (substitution degree 0.73mmol/g) (1648.5 g, 1.0 equiv) and add in solid state reaction still, add DCM(10 L), temperature control 10-30 ℃ is stirred swelling 30min, and suction filtration is removed liquid.Add again DMF(10 L * 2) washing.Measure 20%(v/v, lower same) Piperidine/DMF solution (8.50 L), add in solid state reaction still, temperature control 20-30 ℃ of stirring reaction 5min, suction filtration is removed liquid.Again measure 20%Piperidine/DMF solution (8.50 L), add in solid state reaction still, temperature control 20-30 ℃ of stirring reaction 10min, suction filtration is removed liquid.Measure DMF(10L * 6) add the rear suction filtration of solid state reaction still washing to remove liquid.Separately take Fmoc-Gly-OH(1071.3 g, 3.0 equiv), Cl-HOBt(671.5 g, 3.3 equiv) add dissolution kettle.Measure DMF(4.50 L) add dissolution kettle, stirring is bathed and is cooled to 10-15 ℃ with cryosel after dissolving completely, adds DIC(558 mL, 3.0 equiv), stir temperature control 10-25 ℃ of reaction 5min, then this reaction solution is joined in solid state reaction still, after 2h, suction filtration is removed liquid.Measure DMF(10L * 3) add the rear suction filtration of solid state reaction still washing to remove liquid.Triketohydrindene hydrate detects and is negative.
Step 2
Mpa (Trt)-Tyr (tBu)-Phe-Gln (Trt)-Asn (Trt)-Cys (Trt)-Pro-D-Arg (pbf)-Gly-RinkAmide-AM-Resin's is synthetic
Step 1 product amino-acid resin is sloughed to Fmoc protecting group by the same way.Take Fmoc-AA-OH(3.0 equiv), Cl-HOBt(3.3 equiv) add dissolution kettle.Measure DMF/DCM=1/1 solvent (5 L) and add dissolution kettle, stirring is bathed and is cooled to 5-10 ℃ with cryosel after dissolving completely, adds DIC(3.0 equiv), stir, temperature control 5-25 ℃ of reaction 5min, then joins this reaction solution in solid state reaction still, and after 2h, suction filtration is removed liquid.Measure DMF(12 L * 3) add the rear suction filtration of solid state reaction still washing to remove liquid.Triketohydrindene hydrate detects, as positive in result, repeats coupling once.After all amino acid couplings finish, with DMF(12 L * 6) wash 6 times.
Step 3
Mpa-Tyr (tBu)-Phe-Gln (Trt)-Asn (Trt)-Cys-Pro-D-Arg (pbf)-Gly-RinkAmide-AM-Resin's is synthetic
In step 2 gained peptide resin, add the DCM solution (15 L) containing 1%TFA, stirring reaction 15min, suction filtration is removed liquid.Repeat again 3 times, carry out altogether going for 4 times protection operation.Use again DMF(12 L * 6) wash 6 times.
Step 4
Mpa-Tyr (tBu)-Phe-Gln (Trt)-Asn (Trt)-Cys-Pro-D-Arg (pbf)-Gly-RinkAmide-AM-Resin's (Sulfide bond between Mpa and Cys) is synthetic
Under lucifuge, in step 3 gained peptide resin, add the DMSO solution (15 L) that contains carbamide peroxide (565.6g, 5eq), control 20-30 ℃ of reaction 30min of temperature.Suction filtration is removed liquid, then uses successively DMF(15 L * 3), MeOH(12 L * 2), DCM(20 L * 2), MeOH(12 L * 2) washing resin.Take out Desmopressin peptide resin, after being dried, obtain 4.98 Kg.
Step 5
Mpa-Tyr-Phe-Gln-Asn-Cys-Pro-D-Arg-Gly-NH
2(Sulfide bond between Mpa and Cys's) is synthetic
Measure TFA(47.5 L), PhSMe(2.5 L) join successively in 80 L reactors, stirring and evenly mixing, controls lysate temperature at 15-20 ℃.Step 4 gained peptide resin is all added to cracking still.Control temperature of reaction 25-30 ℃, reaction 2h.Filter, and with TFA(3 L * 2) wash.Merging filtrate, puts to revolve and steams 25% left and right that is concentrated to original volume under evaporimeter.Gained concentrated solution is joined to sedimentation in the methyl tertiary butyl ether of-10～-5 ℃.Centrifugal, washing after 6 times the thick peptide of pasty state, through drying under reduced pressure, obtain thick peptide 1.32 Kg.The thick peptide of gained is dissolved with pure water 40 L, after fully stirring, filter, obtain thick peptide solution.Thick peptide solution is carried out to HPLC quantitative analysis, obtain object peptide approximately 845.5 g, thick peptide HPLC purity 97.87 %, synthesis yield 65.9%.
Yield calculation formula: the thick peptide peak area of the yield=Desmopressin reference substance concentration * Desmopressin reference substance content * Desmopressin * thick peptide amount/(amount of the thick peptide concentration * synthetic of Desmopressin reference substance peak area * Desmopressin * (Desmopressin molecular weight) * 100%
Embodiment 2 yield=0.499 * 86.63% * 138.141 * 130.8/(123.158 * 0.759 * 0.12 * 1069) * 100%=65.1%
Note: the testing conditions of Desmopressin reference substance and the thick peptide of Desmopressin is consistent with sample size, is all: 20 μ l.
Desmopressin reference substance concentration: 0.499 mg/mL
Desmopressin reference substance peak area: 123.158 mAU*min (Fig. 3)
Desmopressin reference substance content: 86.63 %
The thick peptide configuration concentration of Desmopressin: 0.759 mg/mL
The thick peptide peak area of Desmopressin: 138.141 mAU*min.
Claims (5)
1. solid cyclization becomes a method for Desmopressin, and described method steps is as follows:
Step 1, solid phase synthesis Desmopressin linear peptides resin;
Step 2, by the protecting group Trt deprotection on Cys (Trt) and Mpa (Trt);
Step 3, admittedly encircles and obtains cyclic peptide resin with oxygenant;
Step 4, resin is removed in cracking and blocking group obtains Desmopressin.
2. according to the method described in claim 1, it is characterized in that:
Wherein, linear Desmopressin resin described in step 1, structure is as follows:
Mpa(Trt)-Tyr(tBu)-Phe-Gln(Trt)-Asn(Trt)-Cys(Trt)-Pro-D-Arg(pbf)-Gly-NH-Resin
Wherein, deprotection is that the linear Desmopressin resin that uses reagent A and step 1 to synthesize carries out hybrid reaction described in step 2, and the reaction times is 2 hours, and temperature of reaction is 25 ℃; Wherein the compound method of reagent A is as follows: 1mL TFA is joined in 100mL DCM; Wherein, TFA is trifluoroacetic acid, and DCM is methylene dichloride;
Wherein, the oxygenant B that oxidation is used described in step 3 is selected from: carbamide peroxide solution, and its usage quantity is 100mL, and the reaction times is 2 hours, and temperature of reaction is 25 ℃; The compound method of oxygenant B is: the DMSO(dimethyl sulfoxide (DMSO) that the carbamide peroxide of 3.80g is joined to 100mL) in solution, dissolve;
Wherein, the cracking agent C that cracking is used described in step 4 is selected from: thioanisole and trifluoroacetic acid, and its usage quantity is 350mL, and the reaction times is 2 hours, and temperature of reaction is 25 ℃; The compound method of cracking agent C is: by the PhSMe(thioanisole of 35mL) add in the trifluoroacetic acid of 665mL.
3. according to the method described in claim 1, it is characterized in that: oxygenant B is: the solution of carbamide peroxide and DMSO.
4. according to the method described in claim 1, it is characterized in that: phase oxidative has reduced a large amount of waste liquids that bring because of liquid-phase oxidation, accelerated the reaction times simultaneously, belong to technique greenization transformation, improved total recovery.
5. according to the method described in claim 1, it is characterized in that: use carbamide peroxide few as the by product of oxygenant oxidation, purity higher than 97% and yield higher than 65%.
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| CN104761619A (en) * | 2015-01-06 | 2015-07-08 | 苏州天马医药集团天吉生物制药有限公司 | Desmopressin acetate solid phase preparation technology |
| CN107141340A (en) * | 2017-04-18 | 2017-09-08 | 国药集团川抗制药有限公司 | A kind of synthetic method of Terlipressin |
| CN112062813A (en) * | 2019-06-10 | 2020-12-11 | 翰宇药业（武汉）有限公司 | Synthesis method of desmopressin |
| CN114369142A (en) * | 2021-12-31 | 2022-04-19 | 江苏诺泰澳赛诺生物制药股份有限公司 | Method for purifying desmopressin acetate |
| CN116023441A (en) * | 2022-12-29 | 2023-04-28 | 江苏诺泰澳赛诺生物制药股份有限公司 | Method for preparing purified desmopressin sulfoxide impurity |
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