CN102731624B - 一种固相片段法合成比伐卢定的方法 - Google Patents
一种固相片段法合成比伐卢定的方法 Download PDFInfo
- Publication number
- CN102731624B CN102731624B CN201210196621.2A CN201210196621A CN102731624B CN 102731624 B CN102731624 B CN 102731624B CN 201210196621 A CN201210196621 A CN 201210196621A CN 102731624 B CN102731624 B CN 102731624B
- Authority
- CN
- China
- Prior art keywords
- fmoc
- gly
- otbu
- glu
- solid phase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 40
- OIRCOABEOLEUMC-GEJPAHFPSA-N bivalirudin Chemical compound C([C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)CNC(=O)CNC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 OIRCOABEOLEUMC-GEJPAHFPSA-N 0.000 title claims abstract description 32
- 108010055460 bivalirudin Proteins 0.000 title claims abstract description 31
- 229960001500 bivalirudin Drugs 0.000 title claims abstract description 31
- 239000012634 fragment Substances 0.000 title claims abstract description 19
- 239000007790 solid phase Substances 0.000 title claims abstract description 19
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 18
- 238000003786 synthesis reaction Methods 0.000 title claims abstract description 18
- 108010033276 Peptide Fragments Proteins 0.000 claims abstract description 25
- 102000007079 Peptide Fragments Human genes 0.000 claims abstract description 25
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 claims abstract description 10
- 238000005336 cracking Methods 0.000 claims abstract description 8
- 238000010511 deprotection reaction Methods 0.000 claims abstract description 4
- 230000008878 coupling Effects 0.000 claims abstract description 3
- 238000010168 coupling process Methods 0.000 claims abstract description 3
- 238000005859 coupling reaction Methods 0.000 claims abstract description 3
- 125000004213 tert-butoxy group Chemical group [H]C([H])([H])C(O*)(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 25
- 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 claims description 20
- 238000002360 preparation method Methods 0.000 claims description 15
- 239000003153 chemical reaction reagent Substances 0.000 claims description 14
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 claims description 14
- OTKXCALUHMPIGM-FQEVSTJZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-5-[(2-methylpropan-2-yl)oxy]-5-oxopentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 OTKXCALUHMPIGM-FQEVSTJZSA-N 0.000 claims description 12
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 11
- 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 claims description 10
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims description 10
- 238000006467 substitution reaction Methods 0.000 claims description 10
- 230000000694 effects Effects 0.000 claims description 8
- 125000006239 protecting group Chemical group 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 8
- CBPJQFCAFFNICX-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-methylpentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CC(C)C)C(O)=O)C3=CC=CC=C3C2=C1 CBPJQFCAFFNICX-IBGZPJMESA-N 0.000 claims description 7
- JFLSOKIMYBSASW-UHFFFAOYSA-N 1-chloro-2-[chloro(diphenyl)methyl]benzene Chemical compound ClC1=CC=CC=C1C(Cl)(C=1C=CC=CC=1)C1=CC=CC=C1 JFLSOKIMYBSASW-UHFFFAOYSA-N 0.000 claims description 6
- NTFTULBKHJJQAW-HNNXBMFYSA-N 9h-fluoren-9-ylmethyl n-[(2s)-4-methyl-1-oxopentan-2-yl]carbamate Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CC(C)C)C=O)C3=CC=CC=C3C2=C1 NTFTULBKHJJQAW-HNNXBMFYSA-N 0.000 claims description 6
- 229960000549 4-dimethylaminophenol Drugs 0.000 claims description 5
- -1 Fmoc protecting group amino acid Chemical class 0.000 claims description 5
- 230000004913 activation Effects 0.000 claims description 5
- 150000001413 amino acids Chemical class 0.000 claims description 5
- 230000003213 activating effect Effects 0.000 claims description 4
- 239000012429 reaction media Substances 0.000 claims description 4
- SJVFAHZPLIXNDH-JOCHJYFZSA-N (2r)-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-JOCHJYFZSA-N 0.000 claims description 3
- 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 claims description 3
- FODJWPHPWBKDON-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-[(2-methylpropan-2-yl)oxy]-4-oxobutanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 FODJWPHPWBKDON-IBGZPJMESA-N 0.000 claims description 3
- 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 claims description 3
- HNICLNKVURBTKV-NDEPHWFRSA-N (2s)-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-NDEPHWFRSA-N 0.000 claims description 3
- QXVFEIPAZSXRGM-DJJJIMSYSA-N (2s,3s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-methylpentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H]([C@@H](C)CC)C(O)=O)C3=CC=CC=C3C2=C1 QXVFEIPAZSXRGM-DJJJIMSYSA-N 0.000 claims description 3
- 239000003875 Wang resin Substances 0.000 claims description 3
- KJOZJSGOIJQCGA-UHFFFAOYSA-N dichloromethane;2,2,2-trifluoroacetic acid Chemical group ClCCl.OC(=O)C(F)(F)F KJOZJSGOIJQCGA-UHFFFAOYSA-N 0.000 claims description 3
- 150000003053 piperidines Chemical group 0.000 claims description 3
- 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 claims description 2
- 230000002101 lytic effect Effects 0.000 claims description 2
- 239000007791 liquid phase Substances 0.000 abstract description 5
- 238000011031 large-scale manufacturing process Methods 0.000 abstract description 3
- 238000003912 environmental pollution Methods 0.000 abstract description 2
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 24
- 238000006243 chemical reaction Methods 0.000 description 20
- 239000003146 anticoagulant agent Substances 0.000 description 7
- 238000003746 solid phase reaction Methods 0.000 description 6
- 238000010671 solid-state reaction Methods 0.000 description 6
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 229920000669 heparin Polymers 0.000 description 5
- 108090000765 processed proteins & peptides Proteins 0.000 description 5
- 238000002560 therapeutic procedure Methods 0.000 description 5
- FPIRBHDGWMWJEP-UHFFFAOYSA-N 1-hydroxy-7-azabenzotriazole Chemical compound C1=CN=C2N(O)N=NC2=C1 FPIRBHDGWMWJEP-UHFFFAOYSA-N 0.000 description 4
- 239000007821 HATU Substances 0.000 description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 4
- 239000012317 TBTU Substances 0.000 description 4
- CLZISMQKJZCZDN-UHFFFAOYSA-N [benzotriazol-1-yloxy(dimethylamino)methylidene]-dimethylazanium Chemical compound C1=CC=C2N(OC(N(C)C)=[N+](C)C)N=NC2=C1 CLZISMQKJZCZDN-UHFFFAOYSA-N 0.000 description 4
- 229940127219 anticoagulant drug Drugs 0.000 description 4
- 229960002897 heparin Drugs 0.000 description 4
- 238000011282 treatment Methods 0.000 description 4
- 239000002253 acid Substances 0.000 description 3
- 239000012964 benzotriazole Substances 0.000 description 3
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 238000010532 solid phase synthesis reaction Methods 0.000 description 3
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 208000032843 Hemorrhage Diseases 0.000 description 2
- 208000007814 Unstable Angina Diseases 0.000 description 2
- 230000002785 anti-thrombosis Effects 0.000 description 2
- 238000007887 coronary angioplasty Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 208000019622 heart disease Diseases 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N trifluoroacetic acid Substances OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- SWZCTMTWRHEBIN-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-(4-hydroxyphenyl)propanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C1=CC=C(O)C=C1 SWZCTMTWRHEBIN-QFIPXVFZSA-N 0.000 description 1
- SUBDBMMJDZJVOS-UHFFFAOYSA-N 5-methoxy-2-{[(4-methoxy-3,5-dimethylpyridin-2-yl)methyl]sulfinyl}-1H-benzimidazole Chemical compound N=1C2=CC(OC)=CC=C2NC=1S(=O)CC1=NC=C(C)C(OC)=C1C SUBDBMMJDZJVOS-UHFFFAOYSA-N 0.000 description 1
- DBTMQODRSDEGRZ-UHFFFAOYSA-N 9h-fluoren-9-ylmethyl n-(2-oxoethyl)carbamate Chemical compound C1=CC=C2C(COC(=O)NCC=O)C3=CC=CC=C3C2=C1 DBTMQODRSDEGRZ-UHFFFAOYSA-N 0.000 description 1
- 206010002383 Angina Pectoris Diseases 0.000 description 1
- 208000005189 Embolism Diseases 0.000 description 1
- 206010062506 Heparin-induced thrombocytopenia Diseases 0.000 description 1
- NTUPOKHATNSWCY-JYJNAYRXSA-N Phe-Pro-Arg Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)C1=CC=CC=C1 NTUPOKHATNSWCY-JYJNAYRXSA-N 0.000 description 1
- 208000001647 Renal Insufficiency Diseases 0.000 description 1
- 229940122388 Thrombin inhibitor Drugs 0.000 description 1
- 208000001435 Thromboembolism Diseases 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 208000034158 bleeding Diseases 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 239000007822 coupling agent Substances 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- MQYQOVYIJOLTNX-UHFFFAOYSA-N dichloromethane;n,n-dimethylformamide Chemical compound ClCCl.CN(C)C=O MQYQOVYIJOLTNX-UHFFFAOYSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 229940019334 heparin group antithrombotic drug Drugs 0.000 description 1
- 238000000703 high-speed centrifugation Methods 0.000 description 1
- WQPDUTSPKFMPDP-OUMQNGNKSA-N hirudin Chemical group C([C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC(OS(O)(=O)=O)=CC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H]1NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]2CSSC[C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(=O)N[C@H](C(NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N2)=O)CSSC1)C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=2C=CC(O)=CC=2)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)C(C)C)[C@@H](C)O)CSSC1)C(C)C)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 WQPDUTSPKFMPDP-OUMQNGNKSA-N 0.000 description 1
- 229940006607 hirudin Drugs 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 201000006370 kidney failure Diseases 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000003055 low molecular weight heparin Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- LLYKPZOWCPVRPD-UHFFFAOYSA-N n,n-dimethylpyridin-2-amine;n,n-dimethylpyridin-4-amine Chemical compound CN(C)C1=CC=NC=C1.CN(C)C1=CC=CC=N1 LLYKPZOWCPVRPD-UHFFFAOYSA-N 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 239000003087 receptor blocking agent Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000000250 revascularization Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- WROMPOXWARCANT-UHFFFAOYSA-N tfa trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.OC(=O)C(F)(F)F WROMPOXWARCANT-UHFFFAOYSA-N 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 239000003868 thrombin inhibitor Substances 0.000 description 1
- 230000002537 thrombolytic effect Effects 0.000 description 1
- ZGYICYBLPGRURT-UHFFFAOYSA-N tri(propan-2-yl)silicon Chemical compound CC(C)[Si](C(C)C)C(C)C ZGYICYBLPGRURT-UHFFFAOYSA-N 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 239000002699 waste material Substances 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
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明提供一种固相片段法合成比伐卢定的方法。首先用固相逐步法合成一个13肽片段I-Wang Resin，一个N端被Fmoc保护的7肽片段II，二者采用固相片段法偶联合成比伐卢定-Wang Resin；最后经过脱保护、裂解、纯化，冷冻干燥得到纯比伐卢定。该方法相对已有的固相逐步法或液相片段法来说具有工艺操作简单、收率、纯度高、环境污染小、易大规模化生产等优点。
Description
技术领域
本发明涉及一种固相片段法合成多肽的方法，尤其是比伐卢定的固相片段法合成方法。
背景技术
比伐卢定(Bivalirudin)，是一种近年来应用于临床直接的、特异的、可抑性凝血酶抑制剂，其有效抗凝成分为水蛭素衍生物片段，可以直接通过特异性抑制凝血酶活性而发挥抗凝作用，作用可逆而短暂。早期的临床研究显示：比伐卢定抗凝治疗效果确切，且出血事件的发生率较低，和传统肝素抗凝治疗相比使用更为安全。佛罗里达奥克拉心脏病研究所的Bittl自己主持的一项研究显示，在不稳定型心绞痛及梗塞后心绞痛患者行冠状动脉成形术过程中，比伐卢定的效果比肝素好，比伐卢定组患者术后7天的总终点事件(死亡、血管再通术及心肌梗死)发生率少于肝素组(6.2％vs7.9％，P＝0.039)，出血副作用发生率亦更低(3.5％vs 9.3％，P＜0.001)。2000年12月美国食品药物管理局(FDA)批准比伐卢定在美国上市。美国胸科医师学会(ACCP)抗栓和溶栓治疗循证指南和美国心脏学会和心脏病学会2005年于PCI患者抗凝治疗指南指出，比伐卢定克服了肝素、低分子肝素及水蛭素的缺点，具有良好的临床应用前景。可替代肝素用于PCI、不稳定型心绞痛、急性心肌梗死溶栓的治疗，肝素诱导的血小板减少症的抗栓治疗；也可以用于外周动脉介入治疗、心肺移植手术及肾功能不全患者防栓抗栓治疗亦有良好作用，所以可作为普通肝素和IIb/1IIa受体阻滞剂的替代药和辅助药物。
2007年全球销售达2.55亿美元，目前市场应用正在不断扩大。然而，比伐卢定在我国尚未上市，MedicineS公司也没有在亚洲地区的上市计划，这在很大程度上延误了我国心血管医疗事业一尤其是冠状动脉血管成形术的发展。实现比伐卢定的规模化和产业化生产，能够很好地促进比伐卢定药物在我国医疗事业的应用，提高心脏手术的医疗水平。
比伐卢定化学式为：
D-Phe-Pro-Arg-Pro-Gly-Gly-Gly-Gly-Asn-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-OH
目前国内关于比伐卢定合成报道的不多，其中专利号CN200910028793，提供一种比伐卢定的液相合成方法，这个方案主要是通过液相合成三个全保护的片段：N-端全保护的6肽、中段全保护的6肽、C-端全保护的8肽，然后将这三个片段依次在液相中依次缩合得到全保护的比伐卢定。该方案在规模化生产中产生大量的废液，产品也比较难纯化。上述方案工艺比较繁琐，而且产生的杂质多，产品纯度也比较低，不易进行大规模化生产。
发明内容
本发明提供了一种高收率、提成本、反应条件温和、环境污染小、有利于实现产业化的比伐卢定固相片段法合成工艺。
为了实现上述目标，本发明方案主要包括以下步骤：
1)由替代度为0.4～0.9mmol/g的Wang Resin和Fmoc-Leu-OH反应制备Fmoc-Leu-Wang Resin，再按序列依次连接12个含Fmoc保护基氨基酸，得13肽片段I-Wang Resin；
2)由替代度为0.4～0.9mmol/g的Wang Resin和Fmoc-Gly-OH反应制备Fmoc-Gly-Wang Resin，再按序列依次连接6个含Fmoc保护基氨基酸，得7肽片段-2-Chlorotrityl Chloride Resin，裂解得Fmoc保护N端的7肽片段II；
3)13肽片段I-Wang Resin与7肽片段II以固相片段法合成比伐卢定-WangResin，经脱保护，裂解，纯化冷冻干燥得比伐卢定。
上述比伐卢定固相片段法合成工艺，其中，步骤1)所述的Fmoc-Leu-WangResin是由Wang Resin和Fmoc-Leu-OH在以无水DCM或DMF为反应介质，DIC+HOBt+DMAP偶联剂作用下反应制备；固相合成13肽片段I-Wang Resin，其中依次连接的氨基酸为Fmoc-Tyr(tBu)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Pro-OH、Fmoc-Ile-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Phe-OH、Fmoc-Asp(OtBu)-OH、Fmoc-Gly-OH、Fmoc-Asn(Trt)-OH和Fmoc-Gly-OH，反应介质为在DCM或DMF，脱Fmoc保护基采用的试剂为哌啶-DMF，洗涤试剂为无水DMF、DCM和CH3OH，活化试剂为：A+B+C，其中A为HOAt或HOBt，B为HBTU或HATU或TBTU或DIC，C为DIEA。
步骤2)7肽片段II制备，其中Fmoc-Gly-OH和2-Chlorotrityl Chloride Resin在以无水DCM或DMF为反应介质，在DIC+HOBt+DMAP作用下反应制备Fmoc-Gly-2-Chlorotrityl Chloride Resin；其中Fmoc保护的氨基酸为：Fmoc-Gly-OH、Fmoc-Gly-OH、Fmoc-Pro-OH、Fmoc-Pro-OH、Fmoc-Arg(pbf)-OH、Fmoc-Pro-OH、Fmoc-D-Phe-OH。脱Fmoc保护基采用的试剂为哌啶-DMF，洗涤试剂为无水DMF、DCM和CH3OH，活化试剂为：A+B+C，其A为HOAt或HOBt，B为HBTU或HATU或TBTU，C为DIEA或DIC，裂解试剂为TFA-DCM，优选的为5％TFA-DCM。
步骤3)固相片段法合成比伐卢定，其中13肽片段I-Wang Resin为：Fmoc-Gly-Asn(Trt)-Gly-Asp(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ile-Pro-Glu(OtBu)-Glu(OtBu)-Tyr(OtBu)-Leu-Wang Resin；7肽片段II-2-ChlorotritylChloride Resin为：
Fmoc-D-Phe-Pro-Arg(pbf)-Pro-Gly-Gly-Gly-2-Chlorotrityl Chloride Resin，活化试剂为A+B+C，其A为HOAt或HOBt，B为HBT U或HATU或TBTU，C为DIEA或DIC，脱N端保护基实际为哌啶/DMF，裂解及脱侧链保护基试剂为TFA/H2O/TIS。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动性的前提下，还可以根据这些附图获得其他的附图。
图1为本发明13肽片段I-Wang Resin合成工艺流程图；
图2为本发明7肽片段II合成工艺流程图；
图3为本发明固相片段法合成比伐卢定的工艺流程图。
具体实施方式
下面给出的实例以便对本发明进行具体描述，有必要在此指出，本发明不受本方法作为唯一的限制，在本领域对本发明做出一些非本质改动和调整，都属于本发明的保护范围。
在本权利要求书和说明书中出现的缩写代表的意义为下表：
| Fmoc | 9-芴甲氧羰基 |
| HOBt | 1-羟基苯并三唑 |
| HOAt | N-羟基-7-偶氮苯并三氮唑 |
| HATU | 2-(7-偶氮苯并三氮唑)-N，N，N′，N′-四甲基脲六氟磷酸酯 |
| pbf | 2，24，6，7-五甲基-二氢苯并呋喃-5-磺酰基 |
| OtBu | 叔丁氧基 |
| DMF | N，N-二甲基甲酰胺 |
| DCM | 二氯甲烷 |
| DIC | N，N′-二异丙基碳二亚胺 |
| HBTU | 2-(7-偶氮苯并三氮唑)-四甲基脲六氟磷酸酯 |
| TBTU | O-苯并三氮唑-N，N，N′，N′-四甲基脲四氟硼酸 |
| TFA | 三氟乙酸 |
| DMAP | 二甲氨基吡啶 |
| DIEA | N，N-二异丙基乙胺 |
| TIS | 三异丙基硅烷 |
| DIC | N，N-二异丙基碳二亚胺 |
具体实施例说明
实施例1
13肽片段I-Wang Resin的制备
称取1g替代度为0.7mmol/g的Wang Resin，加入到固相反应柱中，加入无水10ml DCM溶胀30min后，用无水DMF洗涤3次(每次1min)，分别将1.4mmol的Fmoc-Leu-OH、HOBt溶于5ml无水DMF，再加入1.4mmol的DIC在0～5℃活化5min后，加入到固相反应柱反应10min，再加入0.2mmol DMAP，在30℃反应5h，反应完毕后分别用无水DMF、CH3OH、DCM、DMF洗涤2、1、1、2次，每次不少于1min，检测替代为0.65mmol/g。用0.1mmol乙酸酐和吡啶封闭树脂上未反应的羟基1h，得Fmoc-Leu-Wang。
依次用10ml体积为25％哌啶-DMF脱Fmoc-Leu-Wang上Fmoc保护基2次后(第一次反应5min，中间用无水DMF洗涤一次，第二次反应15min)，依次用无水DMF、CH3OH，DCM、DMF洗涤2、1、1、2次，每次不少于1min。分别取1.3mmol的Fmoc-Tyr(tBu)-OH、HOBt、HBTU溶于0-5℃的无水DMF，加入0.65mmol的DIEA活化5min后，加入到固相反应柱，在30℃反应2.5h，反应终点以茚三酮法检测为准。重复上述步骤依次偶联Fmoc-Glu(OtBu)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Pro-OH、Fmoc-Ile-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Phe-OH、Fmoc-Asp(OtBu)-OH、Fmoc-Gly-OH、Fmoc-Asn(Trt)-OH、Fmoc-Gly-OH，得13肽片段I-Wang Resin((Fmoc-Gly-Asn(Trt)-Gly-Asp(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ile-Pro-Glu(OtBu)-Glu(OtBu)-Tyr(OtBu)-Leu-Wang Resin)。
实施例2
7肽片段II的制备
称取2g替代度为0.7mmol/g的2-Chlorotrityl Chloride Resin，加入到固相反应柱中，加入无水DCM溶胀30min后，用10ml无水DMF洗涤3次(每次1min)，分别加入2.4mmol的Fmoc-Gly-OH、HOBt、溶于5ml无水DMF中，再加入1.4mmol的DIC在0～5℃活化5min后，加入到固相反应柱反应10min，再加入0.4mmol DMAP，在30℃反应5h，用10ml无水DMF洗涤2次，每次1min，检测替代为0.68mmol/g。用0.2mmol乙酸酐和吡啶封闭树脂上未反应的氨基1h，得Fmoc-Gly-2-Chlorotrityl Chloride Resin。依次用20ml体积为25％哌啶/DMF脱Fmoc-Leu-Wang上Fmoc保护基2次后(第一次反应5min，第二次反应10min，中间用10ml DMF洗涤1次)，依次用无水DMF、CH3OH，DCM、DMF洗涤2、1、1、2次，每次不少于1min。分别取2.4mmol的Fmoc-Gly-OH、HOBt、HBTU溶于0-5℃的无水DMF，加入1mmol DIEA活化5min，加入到固相反应柱，在30℃反应2h，反应终点以茚三酮法检测为准。重复上述步骤依次连接Fmoc-Gly-OH、Fmoc-Gly-OH、Fmoc-Pro-OH、Fmoc-Arg(pbf)-OH、Fmoc-Pro-OH、Fmoc-D-Phe-OH。得7肽片段II-2-Chlorotrityl Chloride Resin。
实施例3
固相片段法合成比伐卢定
取过量1.5倍的7肽片段II，过量2倍的HOBt、HUBT、溶于0-5℃的无水DMF，加入过量1.5倍的DIEA活化5min后，加入到固相合成柱中，在0-30℃反应1-5h(反应终点以茚三酮法检测为准)，无水DMF洗涤3次，得伐卢定-WangResin(D-Phe-Pro-Arg(pbf)-Pro-Gly-Gly-Gly-Gly-Asn(Trt)-Gly-Asp(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ile-Pro-Glu(OtBu)-Glu(OtBu)-Tyr(OtBu)-Leu-Wang Resin)。加入15ml 25％的哌啶-DMF为Fmoc脱保护剂2次后(第一次反应5min，第二次反应10min，中间用10ml DMF洗涤1次)，依次用DMF、DCM、CH3OH洗涤2、1、3次，每次不少于2min；加入15ml体积比为95∶2.5∶2.5的TFA/H2O/TIS作为裂解、侧链保护基脱除试剂反应2-3h，后滤入无水冰乙醚在-5～5℃下静止3～5h，高速离心后经高压液相制备，冷冻干燥的精比伐卢定。其纯度＞99.5％，单杂质＜0.2％，总收率达53％。
本实施例中选取了替代度为0.7mmol/g的Wang Resin与Fmoc-Leu-OH反应制备Fmoc-Leu-Wang Resin；替代度为0.7mmol/g的2-Chlorotrityl Chloride Resin和Fmoc-Gly-OH反应制备Fmoc-Gly-2-Chlorotrityl Chloride Resin；还可以选取替代度为0.4～0.9mmol/g范围的任一Wang Resin和Fmoc-Leu-OH反应制备Fmoc-Leu-Wang Resin；以及替代度为0.4～0.9mmol/g范围的任一2-ChlorotritylChloride Resin和Fmoc-Gly-OH反应制备Fmoc-Gly-2-Chlorotrityl Chloride Resin；均能够实现本发明的技术方案，并取得本发明的所述的技术效果。
以上内容是比伐卢定和成路线最佳优选方案之一，以及对本发明所做的进一步详细说明，但并不能认定本发明的具体实施只限于这些说明，在不脱离本发明构思的前提下，还可以做出若干简单推演及替换，都应当视为本发明保护范围。
Claims (4)
1.一种固相片段法合成比伐卢定的方法，包括以下步骤：
1)由替代度为0.4～0.9mmol/g的Wang Resin和Fmoc-Leu-OH反应制备Fmoc-Leu-Wang Resin，再按序列依次连接12个含Fmoc保护基氨基酸，得13肽片段I-Wang Resin；其中Fmoc-Leu-OH和Wang Resin在DIC+HOBt+DMAP作用下反应制备Fmoc-Leu-Wang Resin；固相逐步合成13肽片段I-Wang Resin，其中Fmoc保护的氨基酸为：Fmoc-Tyr(tBu)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Pro-OH、Fmoc-Ile-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Phe-OH、Fmoc-Asp(OtBu)-OH、Fmoc-Gly-OH、Fmoc-Asn(Trt)-OH和Fmoc-Gly-OH；
2)由替代度为0.4～0.9mmol/g的2-Chlorotrityl Chloride Resin和Fmoc-Gly-OH反应制备Fmoc-Gly-2-Chlorotrityl Chloride Resin，再按序列依次连接剩余的6个含Fmoc保护基氨基酸，得7肽片段II-2-Chlorotrityl Chloride Resin，裂解得Fmoc保护N端的7肽片段II；其中Fmoc-Gly-OH和2-Chlorotrityl ChlorideResin在DIEA作用下反应制备Fmoc-Gly-2-Chlorotrityl Chloride Resin；其中Fmoc保护的氨基酸为：Fmoc-Gly-OH、Fmoc-Gly-OH、Fmoc-Pro-OH、Fmoc-Arg(pbf)-OH、Fmoc-Pro-OH和Fmoc-D-Phe-OH，裂解试剂为TFA-DCM；
3)13肽片段I-Wang Resin与7肽片段II以固相片段法合成比伐卢定-WangResin，其中活化试剂为HOBt、HBTU和DIEA，经脱保护，裂解，纯化冷冻干燥得纯比伐卢定。
2.根据权利要求1所述的一种固相片段法合成比伐卢定的方法，其特征在于，反应介质为无水DMF或DCM，脱Fmoc保护基试剂为哌啶/DMF，洗涤试剂为无水DMF、DCM和CH3OH。
3.根据权利要求1所述的一种固相片段法合成比伐卢定的方法，其特征在于，氨基酸活化温度为1～10℃，偶联、裂解温度为1～30℃。
4.根据权利要求1所述的一种固相片段法合成比伐卢定的方法，其特征在于，13肽片段I-Wang Resin为：
Fmoc-Gly-Asn(Trt)-Gly-Asp(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ile-Pro-Glu(OtBu)-Glu(OtBu)-Tyr(OtBu)-Leu-Wang Resin；
7肽片段II-2-Chlorotrityl Chloride Resin为：
Fmoc-D-Phe-Pro-Arg(pbf)-Pro-Gly-Gly-Gly-2-Chlorotrityl Chloride Resin。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210196621.2A CN102731624B (zh) | 2012-06-14 | 2012-06-14 | 一种固相片段法合成比伐卢定的方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210196621.2A CN102731624B (zh) | 2012-06-14 | 2012-06-14 | 一种固相片段法合成比伐卢定的方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102731624A CN102731624A (zh) | 2012-10-17 |
| CN102731624B true CN102731624B (zh) | 2015-09-23 |
Family
ID=46987996
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210196621.2A Active CN102731624B (zh) | 2012-06-14 | 2012-06-14 | 一种固相片段法合成比伐卢定的方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102731624B (zh) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| USRE46830E1 (en) | 2004-10-19 | 2018-05-08 | Polypeptide Laboratories Holding (Ppl) Ab | Method for solid phase peptide synthesis |
| CN103965293B (zh) * | 2013-02-05 | 2020-02-14 | 深圳信立泰药业股份有限公司 | 一种高纯度比伐卢定及其工业化制备方法 |
| CN103880945B (zh) * | 2013-12-28 | 2016-09-07 | 郑州大明药物科技有限公司 | 制备高纯度胸腺法新的方法 |
| US20170029467A1 (en) * | 2015-07-30 | 2017-02-02 | Ambiopharm, Inc. | Method of producing bivalirudin |
| CN107344967A (zh) * | 2017-06-06 | 2017-11-14 | 牡丹江友搏药业有限责任公司 | 一种比伐卢定的制备及纯化方法 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007033383A2 (en) * | 2005-09-14 | 2007-03-22 | Novetide, Ltd. | Process for production of bivalirudin |
| CN102264757A (zh) * | 2008-12-29 | 2011-11-30 | 隆萨布莱纳公司 | 制备比伐卢定的方法 |
| CN102286076A (zh) * | 2011-06-23 | 2011-12-21 | 成都圣诺科技发展有限公司 | 比伐卢定的制备方法 |
-
2012
- 2012-06-14 CN CN201210196621.2A patent/CN102731624B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007033383A2 (en) * | 2005-09-14 | 2007-03-22 | Novetide, Ltd. | Process for production of bivalirudin |
| CN102264757A (zh) * | 2008-12-29 | 2011-11-30 | 隆萨布莱纳公司 | 制备比伐卢定的方法 |
| CN102286076A (zh) * | 2011-06-23 | 2011-12-21 | 成都圣诺科技发展有限公司 | 比伐卢定的制备方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102731624A (zh) | 2012-10-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102731624B (zh) | 一种固相片段法合成比伐卢定的方法 | |
| CN101033249B (zh) | 固相多肽合成比筏芦定的制备方法 | |
| CN102286076B (zh) | 比伐卢定的制备方法 | |
| FI102183B (fi) | Menetelmä uusien trombiini-inhibiittoreiden valmistamiseksi | |
| CN104031127B (zh) | 一种固液结合制备比伐卢定的方法 | |
| CA2079778C (en) | Inhibitors of thrombin | |
| CN101747426B (zh) | 一种合成普兰林肽的方法 | |
| CN101538316B (zh) | 一种固相法制备Eptifibatide的方法 | |
| Szewczuk et al. | Conformationally restricted thrombin inhibitors resistant to proteolytic digestion | |
| CN101357938B (zh) | 固相多肽合成Exenatide的制备方法 | |
| CN101538324B (zh) | 一种制备艾塞那肽的方法 | |
| CN104974229A (zh) | 一种利那洛肽的固相合成方法 | |
| CN104610433A (zh) | 一种西曲瑞克的制备方法 | |
| CN104418949A (zh) | 一种替度鲁肽的制备方法 | |
| CN104045705A (zh) | 一种利拉鲁肽的合成方法 | |
| CN110204611B (zh) | 一种固相片段法合成比伐卢定 | |
| CN104530224A (zh) | 一种固相合成比伐芦定的方法 | |
| CN103951744A (zh) | 一种固相树脂及其制备方法和用途 | |
| CN109134615B (zh) | 一种比伐芦定的制备方法 | |
| Wishart et al. | A method for the facile solid-phase synthesis of gramicidin S and its analogs | |
| CN106397580A (zh) | 一种比伐卢定的合成方法 | |
| CN109912709B (zh) | 一种酸敏感离子通道抑制剂的制备方法 | |
| CN108383905A (zh) | 一种比伐卢定的制备方法 | |
| CN103319570A (zh) | 一种比伐卢定的制备方法 | |
| CN101033248B (zh) | 一种制备小牛胸腺alpha1的新工艺 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160415 Address after: 224555 Jiangsu Industrial Park, Binhai Economic Development Zone, Yancheng City, Zhongshan Province, two Patentee after: YANCHENG KAILI PHARMACEUTICAL Co.,Ltd. Address before: 214000 Jiangsu city of Wuxi province Yixing Zhoutie town Peng Gan Cun Patentee before: WUXI KAILI PHARMACEUTICAL Co.,Ltd. |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Method for synthesizing bivalirudin by solid phase fragment method Effective date of registration: 20211216 Granted publication date: 20150923 Pledgee: Binhai Haibin Investment Development Co.,Ltd. Pledgor: YANCHENG KAILI PHARMACEUTICAL Co.,Ltd. Registration number: Y2021980015205 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20230626 Granted publication date: 20150923 Pledgee: Binhai Haibin Investment Development Co.,Ltd. Pledgor: YANCHENG KAILI PHARMACEUTICAL Co.,Ltd. Registration number: Y2021980015205 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A Method for the Synthesis of Bivaludine by Solid Phase Fragment Method Effective date of registration: 20230627 Granted publication date: 20150923 Pledgee: Binhai Haibin Investment Development Co.,Ltd. Pledgor: YANCHENG KAILI PHARMACEUTICAL Co.,Ltd. Registration number: Y2023990000319 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20231124 Granted publication date: 20150923 Pledgee: Binhai Haibin Investment Development Co.,Ltd. Pledgor: YANCHENG KAILI PHARMACEUTICAL Co.,Ltd. Registration number: Y2023990000319 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right |