CN111057139B - Novel process for preparing teriparatide - Google Patents
Novel process for preparing teriparatide Download PDFInfo
- Publication number
- CN111057139B CN111057139B CN201811206844.6A CN201811206844A CN111057139B CN 111057139 B CN111057139 B CN 111057139B CN 201811206844 A CN201811206844 A CN 201811206844A CN 111057139 B CN111057139 B CN 111057139B
- Authority
- CN
- China
- Prior art keywords
- fmoc
- trt
- resin
- leu
- fragment
- 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
- 108010049264 Teriparatide Proteins 0.000 title claims abstract description 60
- OGBMKVWORPGQRR-UMXFMPSGSA-N teriparatide Chemical compound C([C@H](NC(=O)[C@H](CCSC)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)[C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CNC=N1 OGBMKVWORPGQRR-UMXFMPSGSA-N 0.000 title claims abstract description 53
- 229960005460 teriparatide Drugs 0.000 title claims abstract description 52
- 238000004519 manufacturing process Methods 0.000 title description 2
- 239000011347 resin Substances 0.000 claims abstract description 88
- 229920005989 resin Polymers 0.000 claims abstract description 88
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 67
- 239000012634 fragment Substances 0.000 claims abstract description 66
- 238000000034 method Methods 0.000 claims abstract description 26
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 claims abstract description 14
- 238000005336 cracking Methods 0.000 claims abstract description 14
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 10
- 238000010189 synthetic method Methods 0.000 claims abstract description 5
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 49
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-dimethylformamide Substances CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 46
- 238000005859 coupling reaction Methods 0.000 claims description 39
- 239000003795 chemical substances by application Substances 0.000 claims description 33
- 150000001413 amino acids Chemical class 0.000 claims description 29
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 28
- 238000006243 chemical reaction Methods 0.000 claims description 21
- 238000010511 deprotection reaction Methods 0.000 claims description 21
- 125000004213 tert-butoxy group Chemical group [H]C([H])([H])C(O*)(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 20
- 230000009471 action Effects 0.000 claims description 19
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 18
- 230000015572 biosynthetic process Effects 0.000 claims description 17
- -1 fmoc-Val-OH Chemical compound 0.000 claims description 17
- 238000003786 synthesis reaction Methods 0.000 claims description 17
- 239000007790 solid phase Substances 0.000 claims description 15
- 238000010532 solid phase synthesis reaction Methods 0.000 claims description 14
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 claims description 12
- 239000011259 mixed solution Substances 0.000 claims description 12
- XXMYDXUIZKNHDT-QNGWXLTQSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-(1-tritylimidazol-4-yl)propanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C(N=C1)=CN1C(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 XXMYDXUIZKNHDT-QNGWXLTQSA-N 0.000 claims description 9
- REITVGIIZHFVGU-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[(2-methylpropan-2-yl)oxy]propanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](COC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 REITVGIIZHFVGU-IBGZPJMESA-N 0.000 claims description 9
- 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 9
- 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 9
- 238000003776 cleavage reaction Methods 0.000 claims description 9
- 230000007017 scission Effects 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 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 8
- 229920001184 polypeptide Polymers 0.000 claims description 8
- 239000000047 product Substances 0.000 claims description 8
- RHQDFWAXVIIEBN-UHFFFAOYSA-N Trifluoroethanol Chemical compound OCC(F)(F)F RHQDFWAXVIIEBN-UHFFFAOYSA-N 0.000 claims description 7
- 239000003875 Wang resin Substances 0.000 claims description 7
- NERFNHBZJXXFGY-UHFFFAOYSA-N [4-[(4-methylphenyl)methoxy]phenyl]methanol Chemical compound C1=CC(C)=CC=C1COC1=CC=C(CO)C=C1 NERFNHBZJXXFGY-UHFFFAOYSA-N 0.000 claims description 7
- 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 claims description 6
- UMRUUWFGLGNQLI-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-6-[(2-methylpropan-2-yl)oxycarbonylamino]hexanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCCCNC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 UMRUUWFGLGNQLI-QFIPXVFZSA-N 0.000 claims description 6
- 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 6
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 6
- 239000012043 crude product Substances 0.000 claims description 6
- BUUKFBVDKSFMHN-LKMAISLMSA-N parathar acetate Chemical compound CC(O)=O.C([C@H](NC(=O)[C@H](CCSC)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)[C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CN=CN1 BUUKFBVDKSFMHN-LKMAISLMSA-N 0.000 claims description 6
- 229960000338 teriparatide acetate Drugs 0.000 claims description 6
- 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 5
- AAAFQLPJNOITCL-SFHVURJKSA-N 9h-fluoren-9-ylmethyl n-[(2s)-1-oxo-3-phenylpropan-2-yl]carbamate Chemical compound C([C@@H](C=O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C1=CC=CC=C1 AAAFQLPJNOITCL-SFHVURJKSA-N 0.000 claims description 5
- 108010016626 Dipeptides Proteins 0.000 claims description 5
- 239000003153 chemical reaction reagent Substances 0.000 claims description 5
- 238000010168 coupling process Methods 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- 230000008878 coupling Effects 0.000 claims description 4
- 238000004108 freeze drying Methods 0.000 claims description 4
- ADOHASQZJSJZBT-SANMLTNESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[1-[(2-methylpropan-2-yl)oxycarbonyl]indol-3-yl]propanoic acid Chemical compound C12=CC=CC=C2N(C(=O)OC(C)(C)C)C=C1C[C@@H](C(O)=O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 ADOHASQZJSJZBT-SANMLTNESA-N 0.000 claims description 3
- UGNIYGNGCNXHTR-SFHVURJKSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-methylbutanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](C(C)C)C(O)=O)C3=CC=CC=C3C2=C1 UGNIYGNGCNXHTR-SFHVURJKSA-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
- BUBGAUHBELNDEW-SFHVURJKSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-methylsulfanylbutanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCSC)C(O)=O)C3=CC=CC=C3C2=C1 BUBGAUHBELNDEW-SFHVURJKSA-N 0.000 claims description 2
- 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 2
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 2
- 239000012351 deprotecting agent Substances 0.000 claims 1
- 125000006239 protecting group Chemical group 0.000 abstract description 17
- 230000009286 beneficial effect Effects 0.000 abstract description 3
- 238000009776 industrial production Methods 0.000 abstract description 3
- 238000000746 purification Methods 0.000 abstract description 3
- 239000000243 solution Substances 0.000 description 24
- 239000012065 filter cake Substances 0.000 description 23
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 22
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 14
- 238000005406 washing Methods 0.000 description 14
- 238000005086 pumping Methods 0.000 description 13
- CMWYAOXYQATXSI-UHFFFAOYSA-N n,n-dimethylformamide;piperidine Chemical compound CN(C)C=O.C1CCNCC1 CMWYAOXYQATXSI-UHFFFAOYSA-N 0.000 description 12
- 238000006467 substitution reaction Methods 0.000 description 12
- 239000000203 mixture Substances 0.000 description 11
- 229910052757 nitrogen Inorganic materials 0.000 description 11
- 238000003746 solid phase reaction Methods 0.000 description 9
- 238000003756 stirring Methods 0.000 description 9
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 6
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 238000001291 vacuum drying Methods 0.000 description 6
- YEDUAINPPJYDJZ-UHFFFAOYSA-N 2-hydroxybenzothiazole Chemical compound C1=CC=C2SC(O)=NC2=C1 YEDUAINPPJYDJZ-UHFFFAOYSA-N 0.000 description 5
- 208000001132 Osteoporosis Diseases 0.000 description 5
- 108090000445 Parathyroid hormone Proteins 0.000 description 5
- 239000000706 filtrate Substances 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
- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 description 4
- 102100036893 Parathyroid hormone Human genes 0.000 description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000012535 impurity Substances 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 238000001308 synthesis method Methods 0.000 description 4
- 230000002194 synthesizing effect Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 125000003277 amino group Chemical group 0.000 description 3
- 230000005587 bubbling Effects 0.000 description 3
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 3
- 238000005520 cutting process Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 125000000524 functional group Chemical group 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000006340 racemization Effects 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 230000008961 swelling Effects 0.000 description 3
- 208000006386 Bone Resorption Diseases 0.000 description 2
- GSNUFIFRDBKVIE-UHFFFAOYSA-N DMF Natural products CC1=CC=C(C)O1 GSNUFIFRDBKVIE-UHFFFAOYSA-N 0.000 description 2
- 206010017076 Fracture Diseases 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- AHVYPIQETPWLSZ-UHFFFAOYSA-N N-methyl-pyrrolidine Natural products CN1CC=CC1 AHVYPIQETPWLSZ-UHFFFAOYSA-N 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000024279 bone resorption Effects 0.000 description 2
- 239000007810 chemical reaction solvent Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 2
- 230000011164 ossification Effects 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 208000001685 postmenopausal osteoporosis Diseases 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000000967 suction filtration Methods 0.000 description 2
- NQRYJNQNLNOLGT-UHFFFAOYSA-N tetrahydropyridine hydrochloride Natural products C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- HBGPNLPABVUVKZ-POTXQNELSA-N (1r,3as,4s,5ar,5br,7r,7ar,11ar,11br,13as,13br)-4,7-dihydroxy-3a,5a,5b,8,8,11a-hexamethyl-1-prop-1-en-2-yl-2,3,4,5,6,7,7a,10,11,11b,12,13,13a,13b-tetradecahydro-1h-cyclopenta[a]chrysen-9-one Chemical compound C([C@@]12C)CC(=O)C(C)(C)[C@@H]1[C@H](O)C[C@]([C@]1(C)C[C@@H]3O)(C)[C@@H]2CC[C@H]1[C@@H]1[C@]3(C)CC[C@H]1C(=C)C HBGPNLPABVUVKZ-POTXQNELSA-N 0.000 description 1
- PFRGGOIBYLYVKM-UHFFFAOYSA-N 15alpha-hydroxylup-20(29)-en-3-one Natural products CC(=C)C1CCC2(C)CC(O)C3(C)C(CCC4C5(C)CCC(=O)C(C)(C)C5CCC34C)C12 PFRGGOIBYLYVKM-UHFFFAOYSA-N 0.000 description 1
- XTWYTFMLZFPYCI-KQYNXXCUSA-N 5'-adenylphosphoric acid Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O XTWYTFMLZFPYCI-KQYNXXCUSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- XTWYTFMLZFPYCI-UHFFFAOYSA-N Adenosine diphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(O)=O)C(O)C1O XTWYTFMLZFPYCI-UHFFFAOYSA-N 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 208000010392 Bone Fractures Diseases 0.000 description 1
- 206010065687 Bone loss Diseases 0.000 description 1
- 101001135732 Bos taurus Parathyroid hormone Proteins 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- ZAFNJMIOTHYJRJ-UHFFFAOYSA-N Diisopropyl ether Chemical compound CC(C)OC(C)C ZAFNJMIOTHYJRJ-UHFFFAOYSA-N 0.000 description 1
- 101001135770 Homo sapiens Parathyroid hormone Proteins 0.000 description 1
- 101001135995 Homo sapiens Probable peptidyl-tRNA hydrolase Proteins 0.000 description 1
- 208000007101 Muscle Cramp Diseases 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 102000003982 Parathyroid hormone Human genes 0.000 description 1
- 102000015731 Peptide Hormones Human genes 0.000 description 1
- 108010038988 Peptide Hormones Proteins 0.000 description 1
- SOKRNBGSNZXYIO-UHFFFAOYSA-N Resinone Natural products CC(=C)C1CCC2(C)C(O)CC3(C)C(CCC4C5(C)CCC(=O)C(C)(C)C5CCC34C)C12 SOKRNBGSNZXYIO-UHFFFAOYSA-N 0.000 description 1
- 229930003316 Vitamin D Natural products 0.000 description 1
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 1
- 239000007825 activation reagent Substances 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 230000037182 bone density Effects 0.000 description 1
- 230000008468 bone growth Effects 0.000 description 1
- 210000001217 buttock Anatomy 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000006317 isomerization reaction Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000004897 n-terminal region Anatomy 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- 210000002997 osteoclast Anatomy 0.000 description 1
- 239000000199 parathyroid hormone Substances 0.000 description 1
- 229960001319 parathyroid hormone Drugs 0.000 description 1
- 239000000813 peptide hormone Substances 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000002953 preparative HPLC Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 206010041569 spinal fracture Diseases 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 235000019166 vitamin D Nutrition 0.000 description 1
- 239000011710 vitamin D Substances 0.000 description 1
- 150000003710 vitamin D derivatives Chemical class 0.000 description 1
- 229940046008 vitamin d Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/635—Parathyroid hormone, i.e. parathormone; Parathyroid hormone-related peptides
-
- 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
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Endocrinology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention provides a synthetic method of teriparatide. In the method, the peptide sequence of the main chain of the teriparatide is divided into a fragment [1-12] peptide resin and a fragment I and a fragment II, wherein the three short peptides can be respectively synthesized first, then the fragment [1-12] peptide resin is connected with the fragment I and the fragment II one by one, the terminal Fmoc protecting group is removed to obtain the teriparatide resin, and finally the target product is obtained through cracking, purification and salification. The structure of each fragment and the fragment resin are described in the specification. The method is simple to operate, and the obtained product has higher purity and lower cost, and is beneficial to industrial production.
Description
Technical Field
The invention relates to a polypeptide synthesis technology, in particular to a synthesis technology for realizing teriparatide by utilizing a solid phase synthesis technology.
Background
Teriparatide Forteo (Teriparatide) was developed for us gift and used for primary osteoporosis, hypogonadal osteoporosis, and postmenopausal osteoporosis. Is a new bone formation promoting drug approved for the first time by the U.S. Food and Drug Administration (FDA), and the parathyroid hormone derivative can promote bone growth by increasing the activity and quantity of osteoblasts, while the conventional osteoporosis drugs at present generally act on the osteoclasts only to slow or block bone loss. The first time teriparatide was marketed in the united states in 12 months 2002 and in china in 3 months 2011. The results of a clinical study involving 1,637 postmenopausal osteoporosis patients showed that 96% of patients showed a significant increase in bone (mineral) density BMD in both the spine and buttocks after treatment with the drug, compared to those who had been administered calcium and vitamin D supplements alone. It has also been found that the agent reduces the risk of developing vertebral fractures and other types of fractures by 65% and 53%, respectively. Another feature of the composition is that it has very little side effects, usually only nausea, dizziness and leg cramps. Teriparatide was sold worldwide in 2016 for $15 billion. China is the country with the largest number of osteoporosis people worldwide, and has data to show that the number of osteoporosis patients in China is 9000 ten thousand at present, and the number is expected to rise to 1.5 hundred million in 2025, so that the market potential is huge.
Teriparatide has the structural formula:
Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly-Lys-His-Leu-Asn-Ser-Met-Glu- Arg-Val-Glu-Trp-Leu-Arg-Lys-Lys-Leu-Gln-Asp-Val-His-Asn-Phe-OH
teriparatide [ hPTH (1-34) ] is a synthetic polypeptide hormone, which is a 1-34 amino acid fragment of human parathyroid hormone PTH, which is the N-terminal region of 84 amino acids containing endogenous parathyroid hormone PTH having biological activity. The immunological and biological properties of teriparatide are identical to those of endogenous parathyroid hormone PTH and bovine parathyroid hormone PTH (bPTH), and the teriparatide can stimulate bone formation and bone resorption, reduce the incidence of fracture in postmenopausal women, and can also improve or reduce bone density according to the different administration modes. Continuous infusion can result in sustained increases in parathyroid hormone PTH concentration and thus greater bone resorption than would occur with daily injections which only result in a brief increase in serum parathyroid hormone PTH concentration. In addition, teriparatide does not inhibit platelet aggregation response of either the adenosine diphosphate-induced pathway or the collagen-induced pathway. The dosage of the medicine is in the range of 31-85 years old, and the increase of the age has no obvious influence on the pharmacokinetics of the medicine and does not need to adjust the dosage.
The patents related to the synthesis of teriparatide at present are as follows:
1) Adopts a step-by-step solid phase synthesis method. According to the amino acid sequence, the CN107501408, CN104530218 and other patents, the peptide resin is obtained by a method of connecting amino acid and resin one by one, then the crude teriparatide is obtained by acidolysis, and the pure teriparatide acetate is obtained by purifying and converting the crude teriparatide into acetate; CN104017064 couples Ser and Met at positions 17 and 18 in order from C-terminus to N-terminus into a dipeptide and then docks into the backbone; CN1103467595 couples Asn and Ser at positions 18 and 19 in order from C-terminus to N-terminus into a dipeptide and then docks into the backbone. Generally CN104017064 and CN1103467595 are also connected in a stepwise manner. In general, the technical method differences of the stepwise solid phase synthesis method are mainly characterized by differences of resin, condensation or activation reagent, cracking solvent and the like, and the advantages of the technical method differences are not obvious.
2) Fragment solid phase synthesis. The amino acid sequences of the CN105384809, the CN102731643 and the like are divided into a plurality of fragments for synthesis, and then the final peptide resin is obtained by splicing, and the yield is 20-40%. The defects are that the number of fragments is large, the steps are long, the yield is low, and improvement and promotion are needed.
In summary, the existing synthetic method of teriparatide has the problems of 20-42% of peptide yield, more impurities, low yield, racemization of amino acid, difficult purification, low yield and the like, and the technical scheme which is suitable for industrial production is not seen. Therefore, the inventor researches on a synthetic method of teriparatide, so that the technical scheme of the invention is obtained.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a teriparatide synthesis method aiming at the problems of more impurities, low purity and yield, high cost, complicated operation steps, excessive waste liquid and adverse industrial production in the existing synthesis process.
The technical problem to be solved by the invention is realized by the following technical scheme, and the synthetic method of teriparatide comprises the following steps:
step one, according to the amino acid sequence from the C end to the N end of a main chain, solid-phase synthesizing fragment [1-12] peptide resin of the 1 st to 12 th amino acids, wherein a protecting group is coupled on a side chain of the peptide resin of the polypeptide fragment;
step two, according to the amino acid sequence from the C end to the N end of the main chain, solid-phase synthesizing peptide resin of 13 th to 22 th amino acids, and cutting the resin to obtain a fragment I;
step three, according to the amino acid sequence from the C end to the N end of the main chain, solid-phase synthesizing the peptide resin of 23 th to 34 th amino acids, and cutting the resin to obtain a fragment II;
step four, removing the tail end Fmoc of the peptide resin of the fragment [1-12] by a solid phase method, and then carrying out coupling reaction on the fragment-I to obtain the peptide resin of the fragment [1-22 ];
step five, removing the tail end Fmoc of the fragment [1-22] peptide resin by a solid phase method, and then carrying out coupling reaction on the fragment II to obtain teriparatide resin;
step six: carrying out a cracking reaction on teriparatide resin under the action of a cracking agent to obtain a crude teriparatide product; step seven: and purifying, salifying and freeze-drying the teriparatide crude product to obtain teriparatide acetate refined peptide.
In the method of the present invention, the peptide sequence of the teriparatide main chain is divided into a short-chain fragment resin and two short-chain fragments. Respectively synthesizing fragment peptide resin and other two fragments, then connecting one by one, and cutting the resin to obtain a crude product of a final product. Numbering the amino acid sequence from the C end to the N end of the teriparatide main chain, wherein the peptide sequence is shown as the following formula:
Ser 34 -Val 33 -Ser 32 -Glu 31 -Ile 30 -Gln 29 -Leu 28 -Met 27 -His 26 -Asn 25 -Leu 24 -Gly 23 -Lys 22 -His 21 -Leu 20 -Asn 19 -Ser 18 -Met 17 -Glu 16 -Arg 15 -Val 14 -Glu 13 -Trp 12 -Leu 11 -Arg 10 -Lys 9 -Lys 8 -Leu 7 -Gln 6 -Asp 5 -Val 4 -His 3 -Asn 2 -Phe 1 -OH
further, the method comprises the steps of,
the segment [1-12] peptide resin, the segment [1-22] peptide resin and His, asp, gln, lys, arg, trp, glu, ser, asn of the teriparatide resin are provided with protecting groups;
the His, asp, gln, lys, arg, trp, glu, ser, asn of the first segment and the second segment are provided with protecting groups;
the protecting group is a protecting group which is commonly used in the field of amino acid synthesis and is used for protecting amino groups on an amino acid main chain and side chain functional groups comprising active hydrogen-containing nitrogen groups or hydroxyl groups or carboxyl groups and other groups interfering synthesis, so that impurities are prevented from being generated in the process of preparing target products by the main chain amino groups or the side chain functional groups and the like.
Preferably, the side chain protecting groups of Asn, gln are Trt;
the side chain protecting group of His is Adoc, boc, mmt, mtt or Trt, further, trt is preferable;
the side chain protecting group of Lys and Trp is Boc or Trt, and further, boc is preferred;
the side chain protecting group of Arg is Boc or Pbf, and further, pbf is preferable;
the side chain protecting group of Glu is preferably OtBu;
the side chain protecting group of Ser is TES, TMS, TBS, DHP, THP or tBu, further preferably tBu.
A solid phase synthesis method of teriparatide, including but not limited to side chain protecting groups and described as preferred protecting groups, as a preferred embodiment, the synthesis method comprises the steps of:
step one, adding Fmoc-Phe-Wang Resin into a solid phase reactor, carrying out coupling reaction with Fmoc-Asn (Trt) -OH under the action of a condensing agent after Fmoc removal reaction, and repeating the deprotection reaction and the coupling reaction, wherein the following amino acids Fmoc-His (Trt) -OH, fmoc-Val-OH, fmoc-Asp (OtBu) -OH, fmoc-Gln (Trt) -OH, fmoc-Leu-OH, fmoc-Lys (Boc) -OH, fmoc-Arg (pbf) -OH, fmoc-Leu-OH and Fmoc-Trp (Boc) -OH are sequentially connected according to a polypeptide sequence to obtain the polypeptide fragment [1-12] peptide Resin;
step two, coupling reaction is carried out on 2-CTC Resin and amino acid Fmoc-Glu (OtBu) -OH under the action of condensing agent to obtain joint Resin, fomc deprotection reaction is carried out on the joint Resin, then adopting a one-by-one solid phase coupling reaction mode to sequentially couple Fmoc-Val-OH Fmoc-Arg (pbf) -OH,
Fmoc-Glu (OtBu) -OH, fmoc-Met-OH, fmoc-Ser (tBu) -OH, fmoc-Asn (Trt) -OH, fmoc-Leu-OH, fmoc-His (Trt) -OH and Fmoc-Lys (Boc) -OH, repeating the coupling reaction and deprotection reaction operations until a fragment-peptide resin is synthesized, and finally, cleaving the resin under the action of a cleavage agent to obtain a fragment-I;
step three, adding the 2-CTC Resin into a solid phase reactor to carry out coupling reaction with amino acid Fmoc-Gly-OH under the action of condensing agent to obtain joint Resin, the preparation method comprises the steps of carrying out Fomc deprotection reaction, and then adopting a one-by-one solid phase coupling reaction mode to sequentially couple amino acid Fmoc-Leu-OH,
Fmoc-Asn(Trt)-OH、Fmoc-His(Trt)-OH、Fmoc-Met-OH、Fmoc-Leu-OH、
Fmoc-Gln (Trt) -OH, fmoc-Ile-OH, fmoc-Glu (OtBu) -OH, fmoc-Ser (tBu) -OH, fmoc-Val-OH and Fmoc-Ser (tBu) -OH, repeating the coupling reaction and deprotection reaction operations until the synthesis to obtain a fragment dipeptide resin, and finally, under the action of a cleavage agent, cleaving the resin to obtain a fragment II;
step four, carrying out deprotection reaction on Fmoc at the tail end of the peptide resin of the fragment [1-12] by a solid phase method, and coupling the Fmoc with the fragment I under the action of a condensing agent to obtain the peptide resin of the fragment [1-22 ];
step five, carrying out deprotection reaction on Fmoc at the tail end of the fragment [1-22] peptide resin by a solid phase method, and coupling the Fmoc with the fragment II under the action of a condensing agent to obtain teriparatide resin;
step six: carrying out a cracking reaction on teriparatide resin under the action of a cracking agent to obtain a crude teriparatide product; step seven: and purifying, salifying and freeze-drying the teriparatide crude product to obtain teriparatide acetate refined peptide.
Further, the method comprises the steps of,
the structural formula of the fragment [1-12] peptide resin is as follows:
Fmoc-Trp 12 (Boc)-Leu 11 -Arg 10 (pbf)-Lys 9 (Boc)-Lys 8 (Boc)-Leu 7 -Gln 6 (Trt)-Asp 5 (OtBu)- Val 4 -His 3 (Trt)-Asn 2 (Trt)-Phe 1 wang resin;
the structural formula of the fragment one is as follows:
Fmoc-Lys 22 (Boc)-His 21 (Trt)-Leu 20 -Asn 19 (Trt)-Ser 18 (tBu)-Met 17 -Glu 16 (OtBu)-Arg 15 (pb f)-Val 14 -Glu 13 (OtBu)-OH；
the structural formula of the segment II is as follows:
Fmoc-Ser 34 (tBu)-Val 33 -Ser 32 (tBu)-Glu 31 (OtBu)-Ile 30 -Gln 29 (Trt)-Leu 28 -Met 27 -His 26 (Trt )-Asn 25 (Trt)-Leu 24 -Gly 23 -OH；
the structural formula of the fragment [1-22] peptide resin is as follows:
Fmoc-Lys 22 (Boc)-His 21 (Trt)-Leu 20 -Asn 19 (Trt)-Ser 18 (tBu)-Met 17 -Glu 16 (OtBu)-Arg 15 (pb f)-Val 14 -Glu 13 (OtBu)-Trp 12 (Boc)-Leu 11 -Arg 10 (pbf)-Lys 9 (Boc)-Lys 8 (Boc)-Leu 7 -Gln 6 (Tr t)-Asp 5 (OtBu)-Val 4 -His 3 (Trt)-Asn 2 (Trt)-Phe 1 wang resin;
the teriparatide resin has the structural formula:
Ser 34 (tBu)-Val 33 -Ser 32 (tBu)-Glu 31 (OtBu)-Ile 30 -Gln 29 (Trt)-Leu 28 -Met 27 -His 26 (Trt)-Asn 25 (Trt)-Leu 24 -Gly 23 -Lys 22 (Boc)-His 21 (Trt)-Leu 20 -Asn 19 (Trt)-Ser 18 (tBu)-Met 17 -Glu 16 (Ot Bu)-Arg 15 (pbf)-Val 14 -Glu 13 (OtBu)-Trp 12 (Boc)-Leu 11 -Arg 10 (pbf)-Lys 9 (Boc)-Lys 8 (Boc)- Leu 7 -Gln 6 (Trt)-Asp 5 (OtBu)-Val 4 -His 3 (Trt)-Asn 2 (Trt)-Phe 1 wang resin.
Further, the method comprises the steps of,
in any of the first to fifth steps, the coupling reaction is performed under the action of a condensing agent, and the condensing agent is selected from one or more of DIC/HOBt, HBTU/HOBt/DIEA, pyBop/HOBt/DIEA, preferably DIC/HOBt; the reaction solvent used is preferably one or more of dichloromethane, DMF, NMP, DMSO, preferably DMF; the deprotection reaction is carried out under the action of a deprotection agent selected from any one of Pip/DMF, naOH/MeOH and TEA/DMF, preferably a solution of Pip/DMF in 20% by volume.
As a preferred embodiment of the present invention,
in any step of the second or third step, the cracking agent is selected from trifluoroacetic acid/dichloromethane mixed solution or trifluoroethanol/dichloromethane mixed solution.
Further, the amount of the mixed solution is as follows: volume ratio TFA: dcm=0.5 to 5:95.0 to 99.5; the mixed solution of trifluoroethanol and dichloromethane has the volume ratio of TFE: dcm=10 to 30: 70-90; further preferred is TFE: dcm=20: 80 (volume ratio); the volume of the cleavage agent is 5 to 20 times the mass of the peptide resin, preferably 8 to 12 times the mass of the peptide resin.
In one scheme, in the step six, the adopted cracking agent is a mixed solution of TFA, phSMe, TIS and water; the volume ratio of the mixed solution is TFA: phSMe: TIS: water=70 to 97:10 to 1:10 to 1:10 to 1; or the adopted cracking agent is a mixed solution of TFA, EDT and water; the volume ratio of the mixed solution is TFA: EDT: water=90 to 95:1 to 5:1 to 5, the ratio is preferably 90:5:5, a step of;
further, in the step six, the volume of the cracking agent is 5 to 25 times of the mass of the peptide resin, preferably 8 to 12 times of the mass of the peptide resin;
in one of the embodiments of the present invention,
in the solid phase synthesis method of teriparatide, the solid phase carrier for solid phase synthesis is Wang resin with the substitution degree of 0.2-1.0mmol/g, fmoc-Phe-Wang resin with the substitution degree of 0.2-0.7mmol/g, HMP resin with the substitution degree of 0.2-1.0mmol/g, HMBA-AM resin with the substitution degree of 0.2-1.0mmol/g or 2-CTC resin with the substitution degree of 0.2-1.5 mmol/g; fmoc-Phe-Wang resin with a degree of substitution of 0.3-0.5mmol/g or 2-CTC resin with a degree of substitution of 0.6-1.2mmol/g is preferred.
In one of the embodiments of the present invention,
the reaction solvent used in the invention is any one or more selected from dichloromethane and DMF, NMP, DMSO.
In the invention, the mode of solid-phase coupling reaction one by one is usually that a reaction substrate is sequentially connected with Fmoc-protected amino acids, after each coupling reaction is finished, fmoc-protecting group removal reaction is carried out by using a deprotection agent, then solid-phase coupling reaction is carried out with the next Fmoc-protected amino acid, and the coupling reaction and the deprotection reaction operation are sequentially repeated until a product of a target peptide sequence structure is synthesized.
The side chain protecting group is a protecting group which is commonly used in the field of amino acid synthesis and is used for protecting functional groups such as nitrogen groups, hydroxyl groups or carboxyl groups containing hydrogen on a main chain and a side chain of the amino acid, and is helpful for preventing part of the side chain from reacting with chemical substances used in the steps of peptide synthesis, processing and the like.
The condensing agent is a reagent capable of causing a coupling reaction, and particularly a reagent capable of promoting amino groups and carboxyl groups to be coupled to form peptide bonds in polypeptide synthesis.
The activator refers to an agent which can assist a condensing agent to better promote a coupling reaction in the polypeptide coupling reaction, such as: inhibit the generation of racemization impurities in the coupling reaction, accelerate the reaction speed by catalysis, and the like.
In the method of the present invention, the three short peptides can be synthesized first according to the peptide sequence of the teriparatide main chain, which is divided into one basic fragment [1-12] peptide resin and two fragments as described above. And then connecting the fragment [1-12] peptide resin with the fragment I and the fragment II one by one, and removing the terminal Fmoc protecting group to obtain the teriparatide resin. Finally obtaining the target product through cracking, purifying and salifying.
FIG. 1 is a flow chart for the preparation of teriparatide
The beneficial effects of the invention are as follows:
1) The invention adopts three fragment coupling methods to carry out solid phase synthesis of teriparatide, wherein the fragment [1-12] peptide resin, the fragment I and the fragment II are easy to synthesize and purify, and the purity is high;
2) Compared with the step 34 of the stepwise synthesis method, the method shortens the step 13, and different fragments can be synthesized simultaneously, so that the synthetic time of teriparatide is effectively shortened;
3) The fragment synthesis method adopted by the invention has short steps and mild reaction conditions, and can effectively reduce the problems of more amino acid sequence differences, isomerization, racemization of amino acids and the like of the missing peptide and the main component;
4) The product is easier to purify, the post-treatment operation is simple, and the three wastes are reduced;
5) The method has stable process, the final teriparatide purity can reach more than 99.5 percent, the total yield is 46 percent, and the method is beneficial to industrial application.
Detailed Description
The present invention will be described in detail below with reference to specific examples for the purpose of understanding. It is specifically noted that the specific examples are for illustrative purposes only and that modifications are possible within the scope of the invention, as those skilled in the art will readily recognize from the description herein.
Abbreviations or english full names used in the specification and claims of the present invention have the following meanings:
embodiment one: synthesis of fragment [1-12] peptide resins
Fmoc-Phe-Wang resin (10.00 g substitution degree 0.35 mmol/g) was added to the solid phase flask, and the flask was swelled with 200 mM LDCM and then dried. The filter cake was mixed with 300ml of 20% piperidine/DMF solution and was pumped down after stirring for 5 minutes; then 300mL of 20% piperidine/DMF solution was added, mixed and stirred for 15 minutes and then drained. Washing the filter cake with a proper amount of DMF, pumping to dryness, and placing the filter cake in a solid phase reaction bottle for later use. Fmoc-Asn (Trt) -OH (6.45 g), HOBT (1.46 g) and 30mLDMF were sequentially added to another dry reaction flask, stirred, DIC (1.36 g) was added under ice bath for 30min, and stirred to obtain an activated solution. Adding the newly obtained solution into the solid phase reaction bottle, stirring for 2h under the protection of nitrogen until the coupling reaction is detected to be complete by ninhydrin. Pumping, washing the filter cake with a proper amount of DMF, and pumping. The deprotection and coupling reaction steps are repeated, and Fmoc-His (Trt) -OH, fmoc-Val-OH, fmoc-Asp (OtBu) -OH, fmoc-Gln (Trt) -OH, fmoc-Leu-OH, fmoc-Lys (Boc) -OH, fmoc-Arg (pbf) -OH, fmoc-Leu-OH, and Fmoc-Trp (Boc) -OH are sequentially connected. After the coupling reaction is finished, the mixture is pumped out, and the filter cake is washed by DMF, and is alternately washed by MeOH and DCM, and pumped out. Vacuum drying to obtain fragment [1-12] peptide resin, which is directly used in the next step.
0.10g of the peptide resin was taken and put into 1mL of a lysate (TFA: EDT: water, 90:5:5 by volume ratio) and stirred at room temperature for 3 hours. The resin was washed with small amounts of TFA, the filtrates were combined, mixed with 15mL of glacial diethyl ether, settled, centrifuged, and the white precipitate washed with diethyl ether and dried. The solid was obtained by vacuum drying, and the purity was 95% by detection.
Example two Synthesis of fragment one
2-CTC Resin (25.00 g, substitution degree 1.12 mmol/g) and 400mLDCM were added into a solid phase reaction flask in sequence, and suction filtration was performed after swelling to obtain a filter cake. Fmoc-Glu (OtBu) -OH (8.51 g), 100mL DCM were added sequentially, followed by stirring under nitrogen and DIEA (about 9.53 g) was added dropwise. After the dripping, stirring and reacting for 2 hours. MeOH (25 mL) was added and stirring continued for 30min. And (3) pumping, alternately washing a filter cake by MeOH and DCM, pumping, and vacuum drying at room temperature to obtain the joint resin. Detecting the substitution degree of 0.73mmol/g;
the resin and 200mLDCM are added into a solid-phase reaction bottle in sequence, and the mixture is pumped out after swelling. 100mL of 20% piperidine/DMF solution is added, mixed and stirred for 5 minutes, and pumped down; 100mL of 20% piperidine/DMF solution was added, mixed and stirred for 15 minutes, and then drained. Washing the filter cake with a proper amount of DMF and pumping. Fmoc-Val-OH (18.32 g), HOBT (8.75 g), and 100 mM LDMF were placed in another dry reaction flask, stirred, and DIC (8.18 g) was added under nitrogen protection in an ice bath to give an activated solution. Adding the new solution into the solid phase reaction bottle under the protection of nitrogen, and stirring for 2 hours until the ninhydrin detection coupling reaction is complete. The mixture was drained and the filter cake was washed with DMF. Repeating the deprotection and coupling reaction steps, and sequentially connecting Fmoc-Arg (pbf) -OH,
Fmoc-Glu (OtBu) -OH, fmoc-Met-OH, fmoc-Ser (tBu) -OH, fmoc-Asn (Trt) -OH, fmoc-Leu-OH, fmoc-His (Trt) -OH, fmoc-Lys (Boc) -OH. After the coupling reaction is finished, pumping to dryness, washing a filter cake by DMF, alternately washing by MeOH and DCM, pumping to dryness, and drying in vacuum at room temperature to obtain the fragment-peptide resin.
The peptide resin, 500ml of 20% TFE/DCM and nitrogen are added into a reaction flask in sequence, the mixture is stirred for 2 hours under nitrogen bubbling, the mixture is filtered under suction, a proper amount of DCM is used for washing filter cakes, the filtrate is collected, and the filtrate is concentrated under reduced pressure at 30 ℃ to remove the solvent. 200ml of LDCM was added to the residue, dissolved, added with glacial ethyl ether, crystallized, centrifuged, washed with ethyl ether, pumped down, and dried at room temperature to give 41.23g of fragment I with a yield of 91% and a purity of 89%.
Embodiment III: synthesis of fragment two
2-CTC Resin (25.00 g, substitution degree 1.12 mmol/g) and 400 mM LDCM were added into a solid phase reaction flask in sequence, and the mixture was dried after swelling to obtain a filter cake. 100 mM LDCM, fmoc-Gly-OH (5.95 g) and DIEA (9.53 g) were added dropwise with stirring under nitrogen. After the completion of the dropwise addition, the reaction was stirred for 2 hours, meOH (25 mL) was added, and the reaction was stirred for 30 minutes with nitrogen bubbling. The mixture was drained and the filter cake was washed alternately with DCM and MeOH. And (5) pumping, and vacuum drying at room temperature to obtain the joint resin. The substitution degree is 0.74mmol/g through detection.
200mL of DCM swells the above materials and is drained. The filter cake was mixed with 100mL of 20% piperidine/DMF solution and stirred for 5 minutes and drained; 100mL of 20% piperidine/DMF solution was added and mixed and stirred for 15 minutes and then drained. Washing the filter cake with a proper amount of DMF and pumping. 400mL of LDMF was added to the flask and stirred, and Fmoc-Leu-OH (19.08 g), HOBT (8.75 g) and DIC (8.18 g) were added sequentially over 30min in an ice bath. Stirring and reacting for 2h under the protection of nitrogen until the ninhydrin detection reaction is complete. The peptide resin was dried by suction and washed with DMF. The deprotection and coupling reaction steps were repeated and Fmoc-Asn (Trt) -OH, fmoc-His (Trt) -OH, fmoc-Met-OH, fmoc-Leu-OH, fmoc-Gln (Trt) -OH, fmoc-Ile-OH, fmoc-Glu (OtBu) -OH, fmoc-Ser (tBu) -OH, fmoc-Val-OH, fmoc-Ser (tBu) -OH were sequentially connected. After the coupling reaction is finished, the mixture is pumped out, the filter cake is washed by a proper amount of DMF, and is alternately washed by a proper amount of DCM and MeOH, and the mixture is pumped out. Vacuum drying at room temperature to obtain the fragment dipeptide resin.
To the above peptide resin was added 500ml of 20% TFE/DCM, stirred under nitrogen for 2 hours, suction filtration, washing the filter cake with a proper amount of DCM, collecting the filtrate, concentrating under reduced pressure at room temperature to remove the solvent, dissolving the residue with 200ml of LDCM, adding into glacial ethyl ether for crystallization, centrifuging, washing with ethyl ether, pumping, drying at room temperature to obtain 40.96g of fragment two, yield 95%, purity 90%.
Embodiment four: synthesis of fragment [1-22] peptide resins
The peptide resin obtained in example one (13.65 g), 200ml of LDCM was swollen in a solid-phase reaction flask and dried. The filter cake was mixed with 100mL of 20% piperidine/DMF solution and stirred for 5 minutes and drained; 100mL of 20% piperidine/DMF solution was added, mixed and stirred for 15 minutes, and then drained. Washing the filter cake by using a proper amount of DMF, and pumping for later use. In another dry reaction flask, fragment one (9.65 g), HOBT (2.96 g), DMF (50 mL) and DIC (3.00 g) were added sequentially, mixed and activated to obtain a solution, and then added to the solid phase reaction flask. The reaction was carried out under nitrogen bubbling until the ninhydrin detection reaction was complete. 100mL of 30% acetic anhydride/pyridine/DCM solution was added and stirred for 1h and drained. The filter cake was washed with a suitable amount of DMF and pumped dry for direct use in the next step.
Fifth embodiment: synthesis of teriparatide resin
The peptide resin obtained in example four was swelled at 200ml cm and then dried. The filter cake was mixed with 100mL of 20% piperidine/DMF solution and stirred for 5 minutes and drained; 100mL of 20% piperidine/DMF solution was added and the mixture was stirred for 15 minutes and drained. Washing with a proper amount of DMF, and pumping for later use. In another dry reaction flask, fragment two (13.26 g), HOBT (2.96 g), DMF (50 mL) and DIC (3.00 g) were added in this order, activated by stirring to give a solution, and then added to the solid phase reaction flask. The reaction is completed until ninhydrin detection reaction is completed. 100mL of 30% acetic anhydride/pyridine/DCM solution was added and stirred for 1h and then drained. Washing with a proper amount of DMF and draining. The filter cake was mixed with 100mL of 20% piperidine/DMF solution and stirred for 5 minutes and drained; 100mL of 20% piperidine/DMF solution was added, mixed and stirred for 15 minutes, and then drained. The filter cake was washed with a suitable amount of DMF, alternately with a suitable amount of DCM and MeOH, drained and dried in vacuo to give 25.00g of peptide resin which was used directly in the next step.
Example six: synthesis of crude teriparatide
The crude resin obtained in example five was mixed with 150mL of cleavage agent (TFA: water: EDT,90:5:5, v/v) and cleaved at 25℃for 3 hours to the end of the reaction. Filtering, adding appropriate amount of isopropyl ether into the filtrate, settling, centrifuging, washing, separating, and vacuum drying at room temperature to obtain 12.25g of teriparatide crude product with HPLC purity of 75% and total yield of 85%.
Embodiment seven: preparation of teriparatide acetate
Three drops of ammonia water and 1.00g of teriparatide crude product are sequentially added into water, and the solution is dissolved by shaking and purified by a Hanbang preparation HPLC system (the wavelength is 210nm, the chromatographic column is a C8 reverse phase column, the pH is 3.0/acetonitrile is a mobile phase), so that the teriparatide refined peptide solution with the purity of more than 99.5 percent is obtained. Transferring the refined peptide solution into salt by adopting a preparative HPLC system (chromatographic column is C18 reverse phase column, 0.2% acetic acid/acetonitrile is mobile phase), collecting target components, concentrating under reduced pressure, and freeze-drying to obtain 0.55g of teriparatide acetate pure product with purity of 99.7%, purification yield of 55% and total yield of 46%; mass spectrum (M+H) + ＝1373.5,Calculated：4117.5。
Claims (7)
1. The synthetic method of teriparatide is characterized by comprising the following steps:
step one, adding Fmoc-Phe-Wang Resin into a solid phase reactor, carrying out coupling reaction with Fmoc-Asn (Trt) -OH under the action of a condensing agent after Fmoc removal reaction, and repeating the deprotection reaction and the coupling reaction, wherein the following amino acids Fmoc-His (Trt) -OH, fmoc-Val-OH, fmoc-Asp (OtBu) -OH, fmoc-Gln (Trt) -OH, fmoc-Leu-OH, fmoc-Lys (Boc) -OH, fmoc-Arg (pbf) -OH, fmoc-Leu-OH and Fmoc-Trp (Boc) -OH are sequentially connected according to a polypeptide sequence to obtain the polypeptide fragment [1-12] peptide Resin;
step two, coupling reaction is carried out on 2-CTC Resin and amino acid Fmoc-Glu (OtBu) -OH under the action of condensing agent to obtain joint Resin, fomc deprotection reaction is carried out on the joint Resin, then adopting a one-by-one solid phase coupling reaction mode to sequentially couple Fmoc-Val-OH Fmoc-Arg (pbf) -OH,
Fmoc-Glu (OtBu) -OH, fmoc-Met-OH, fmoc-Ser (tBu) -OH, fmoc-Asn (Trt) -OH, fmoc-Leu-OH, fmoc-His (Trt) -OH and Fmoc-Lys (Boc) -OH, repeating the coupling reaction and deprotection reaction operations until a fragment-peptide resin is synthesized, and finally, cleaving the resin under the action of a cleavage agent to obtain a fragment-I;
step three, adding the 2-CTC Resin into a solid phase reactor to carry out coupling reaction with amino acid Fmoc-Gly-OH under the action of condensing agent to obtain joint Resin, the preparation method comprises the steps of carrying out Fomc deprotection reaction, and then adopting a one-by-one solid phase coupling reaction mode to sequentially couple amino acid Fmoc-Leu-OH,
Fmoc-Asn(Trt)-OH、Fmoc-His(Trt)-OH、Fmoc-Met-OH、Fmoc-Leu-OH、
Fmoc-Gln (Trt) -OH, fmoc-Ile-OH, fmoc-Glu (OtBu) -OH, fmoc-Ser (tBu) -OH, fmoc-Val-OH and Fmoc-Ser (tBu) -OH, repeating the coupling reaction and deprotection reaction operations until the synthesis to obtain a fragment dipeptide resin, and finally, under the action of a cleavage agent, cleaving the resin to obtain a fragment II;
step four, carrying out deprotection reaction on Fmoc at the tail end of the peptide resin of the fragment [1-12] by a solid phase method, and coupling the Fmoc with the fragment I under the action of a condensing agent to obtain the peptide resin of the fragment [1-22 ];
step five, carrying out deprotection reaction on Fmoc at the tail end of the fragment [1-22] peptide resin by a solid phase method, and coupling the Fmoc with the fragment II under the action of a condensing agent to obtain teriparatide resin;
step six: carrying out a cracking reaction on teriparatide resin under the action of a cracking reagent to obtain a crude teriparatide product;
step seven: and purifying, salifying and freeze-drying the teriparatide crude product to obtain teriparatide acetate refined peptide.
2. The method according to claim 1, wherein the fragments [1-12]]The peptide resin has the structural formula: fmoc-Trp 12 (Boc)-Leu 11 -Arg 10 (pbf)-Lys 9 (Boc)-Lys 8 (Boc)-Leu 7 -Gln 6 (Trt)-Asp 5 (OtBu)-Val 4 -His 3 (Trt)-Asn 2 (Trt)-Phe 1 Wang resin;
the structural formula of the fragment one is as follows:
Fmoc-Lys 22 (Boc)-His 21 (Trt)-Leu 20 -Asn 19 (Trt)-Ser 18 (tBu)-Met 17 -Glu 16 (OtBu)-Arg 15 (pb f)-Val 14 -Glu 13 (OtBu)-OH；
the structural formula of the segment II is as follows:
Fmoc-Ser 34 (tBu)-Val 33 -Ser 32 (tBu)-Glu 31 (OtBu)-Ile 30 -Gln 29 (Trt)-Leu 28 -Met 27 -His 26 (Trt)-Asn 25 (Trt)-Leu 24 -Gly 23 -OH；
the structural formula of the fragment [1-22] peptide resin is as follows:
Fmoc-Lys 22 (Boc)-His 21 (Trt)-Leu 20 -Asn 19 (Trt)-Ser 18 (tBu)-Met 17 -Glu 16 (OtBu)-Arg 15 (pb f)-Val 14 -Glu 13 (OtBu)-Trp 12 (Boc)-Leu 11 -Arg 10 (pbf)-Lys 9 (Boc)-Lys 8 (Boc)-Leu 7 -Gln 6 (Tr t)-Asp 5 (OtBu)-Val 4 -His 3 (Trt)-Asn 2 (Trt)-Phe 1 wang resin;
the teriparatide resin has the structural formula:
Ser 34 (tBu)-Val 33 -Ser 32 (tBu)-Glu 31 (OtBu)-Ile 30 -Gln 29 (Trt)-Leu 28 -Met 27 -His 26 (Trt)-Asn 25 (Trt)-Leu 24 -Gly 23 -Lys 22 (Boc)-His 21 (Trt)-Leu 20 -Asn 19 (Trt)-Ser 18 (tBu)-Met 17 -Glu 16 (Ot Bu)-Arg 15 (pbf)-Val 14 -Glu 13 (OtBu)-Trp 12 (Boc)-Leu 11 -Arg 10 (pbf)-Lys 9 (Boc)-Lys 8 (Boc)-Leu 7 -Gln 6 (Trt)-Asp 5 (OtBu)-Val 4 -His 3 (Trt)-Asn 2 (Trt)-Phe 1 wang resin.
3. The method according to claim 1, wherein the condensing agent is selected from one or more of DIC/HOBt, HBTU/HOBt/DIEA, pyBop/HOBt/DIEA.
4. The method according to claim 1, wherein the deprotecting agent is selected from any of Pip/DMF, naOH/MeOH, TEA/DMF in any of the steps one to five.
5. The method according to claim 1, wherein in any of the second or third steps, the cleavage agent is selected from the group consisting of trifluoroacetic acid/dichloromethane mixed solution and trifluoroethanol/dichloromethane mixed solution.
6. The method according to claim 5, wherein the volume of the cleavage agent is 5 to 20 times the mass of the peptide resin.
7. The method according to claim 1, wherein in the sixth step, the cleavage reagent is a mixed solution of trifluoroacetic acid; the volume ratio of the trifluoroacetic acid mixed solution is TFA: phSMe: TIS: water=70 to 97:10 to 1:10 to 1:10 to 1; or the volume ratio is TFA: EDT: water=90 to 95:1 to 5:1 to 5; the volume of the cracking reagent is 5-25 times of the mass of the peptide resin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811206844.6A CN111057139B (en) | 2018-10-17 | 2018-10-17 | Novel process for preparing teriparatide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811206844.6A CN111057139B (en) | 2018-10-17 | 2018-10-17 | Novel process for preparing teriparatide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111057139A CN111057139A (en) | 2020-04-24 |
| CN111057139B true CN111057139B (en) | 2023-12-22 |
Family
ID=70296832
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811206844.6A Active CN111057139B (en) | 2018-10-17 | 2018-10-17 | Novel process for preparing teriparatide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111057139B (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4203040A1 (en) * | 1992-02-04 | 1993-08-05 | Boehringer Mannheim Gmbh | NEW PARATHORMON FRAGMENTS, THEIR PRODUCTION AND MEDICINAL PRODUCTS CONTAINING THEM |
| US6849710B1 (en) * | 1996-07-30 | 2005-02-01 | F. Hoffmann-La Roche Ag | Method for the synthesis of analogs of parathyroid hormone and parathyroid hormone related peptide |
| CN102731643A (en) * | 2012-06-26 | 2012-10-17 | 深圳翰宇药业股份有限公司 | Method for preparing polypeptide used for treating osteoporosis |
| CN104017064A (en) * | 2014-06-13 | 2014-09-03 | 杭州诺泰制药技术有限公司 | Method for preparing teriparatide |
| CN104530218A (en) * | 2015-01-07 | 2015-04-22 | 哈尔滨吉象隆生物技术有限公司 | Solid-phase synthesis method of teriparatide |
| CN104910269A (en) * | 2015-06-02 | 2015-09-16 | 成都圣诺生物科技股份有限公司 | Method for synthesizing teriparatide |
| CN105384809A (en) * | 2015-12-30 | 2016-03-09 | 济南康和医药科技有限公司 | Method for preparing teriparatide by fragment method and solid-liquid combination |
| CN107501408A (en) * | 2017-09-22 | 2017-12-22 | 扬子江药业集团四川海蓉药业有限公司 | A kind of preparation method of Teriparatide |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9975941B2 (en) * | 2015-11-11 | 2018-05-22 | Richter-Helm Bio Tec Gmbh & Co. Kg | Method for purifying teriparatide |
-
2018
- 2018-10-17 CN CN201811206844.6A patent/CN111057139B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4203040A1 (en) * | 1992-02-04 | 1993-08-05 | Boehringer Mannheim Gmbh | NEW PARATHORMON FRAGMENTS, THEIR PRODUCTION AND MEDICINAL PRODUCTS CONTAINING THEM |
| US6849710B1 (en) * | 1996-07-30 | 2005-02-01 | F. Hoffmann-La Roche Ag | Method for the synthesis of analogs of parathyroid hormone and parathyroid hormone related peptide |
| CN102731643A (en) * | 2012-06-26 | 2012-10-17 | 深圳翰宇药业股份有限公司 | Method for preparing polypeptide used for treating osteoporosis |
| CN104017064A (en) * | 2014-06-13 | 2014-09-03 | 杭州诺泰制药技术有限公司 | Method for preparing teriparatide |
| CN104530218A (en) * | 2015-01-07 | 2015-04-22 | 哈尔滨吉象隆生物技术有限公司 | Solid-phase synthesis method of teriparatide |
| CN104910269A (en) * | 2015-06-02 | 2015-09-16 | 成都圣诺生物科技股份有限公司 | Method for synthesizing teriparatide |
| CN105384809A (en) * | 2015-12-30 | 2016-03-09 | 济南康和医药科技有限公司 | Method for preparing teriparatide by fragment method and solid-liquid combination |
| CN107501408A (en) * | 2017-09-22 | 2017-12-22 | 扬子江药业集团四川海蓉药业有限公司 | A kind of preparation method of Teriparatide |
Non-Patent Citations (1)
| Title |
|---|
| 周建华等.片段法合成抗骨质疏松多肽药物特立帕肽.合成化学.2018,第26卷(第8期),摘要、第1.2节、第2.1-2.2节. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111057139A (en) | 2020-04-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106699871B (en) | Preparation method of liraglutide | |
| WO2018032521A1 (en) | Method for synthesizing liraglutide | |
| CN106146648B (en) | Synthetic method of parathyroid hormone analogue | |
| CN111087462B (en) | Solid-phase synthesis method of abamectin | |
| CN110294800B (en) | Preparation method of somaglutide | |
| CN103497245B (en) | Method for synthesizing thymalfasin | |
| CN107880111B (en) | Method for preparing liraglutide | |
| CN111732651B (en) | Method for preparing Somalutide through continuous flow solid phase reaction | |
| CN113135991B (en) | Method for preparing somaglutide | |
| CN111732649B (en) | Preparation of liraglutide by continuous flow solid phase reaction | |
| CN113801197A (en) | Preparation method of procatide | |
| WO2020199461A1 (en) | Method for synthesizing polypeptide-derived compound | |
| CN110903352A (en) | Preparation method of cetrorelix | |
| CN112679602A (en) | Solid-phase synthesis method of Somaloutide | |
| CN117106055A (en) | Synthesis method of telipopeptide | |
| CN110615836B (en) | Solid-phase synthesis method of liraglutide | |
| CN111057139B (en) | Novel process for preparing teriparatide | |
| CN110894227A (en) | Solid-phase synthesis method of liraglutide | |
| CN111944040B (en) | Solid phase synthesis method of abamectin | |
| CN115677827A (en) | Peptide compounds | |
| CN112175067B (en) | Preparation method of teduglutide | |
| CN111233980B (en) | Fragment method synthesis method of goserelin | |
| CN113637064B (en) | Synthetic method of somalupeptide | |
| CN103936848B (en) | Thymosin α1Method of synthesis of | |
| CN110845600B (en) | Method for preparing liraglutide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |