CN116874585B - Synthesis method of insulin detention - Google Patents
Synthesis method of insulin detention Download PDFInfo
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- CN116874585B CN116874585B CN202311140918.1A CN202311140918A CN116874585B CN 116874585 B CN116874585 B CN 116874585B CN 202311140918 A CN202311140918 A CN 202311140918A CN 116874585 B CN116874585 B CN 116874585B
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- Prior art keywords
- insulin
- side chain
- nitroimidazole
- detention
- amount
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- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 title claims abstract description 265
- 102000004877 Insulin Human genes 0.000 title claims abstract description 131
- 108090001061 Insulin Proteins 0.000 title claims abstract description 131
- 229940125396 insulin Drugs 0.000 title claims abstract description 131
- 238000001308 synthesis method Methods 0.000 title abstract description 14
- YZEUHQHUFTYLPH-UHFFFAOYSA-N 2-nitroimidazole Chemical compound [O-][N+](=O)C1=NC=CN1 YZEUHQHUFTYLPH-UHFFFAOYSA-N 0.000 claims abstract description 67
- TUNFSRHWOTWDNC-UHFFFAOYSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCCC(O)=O TUNFSRHWOTWDNC-UHFFFAOYSA-N 0.000 claims abstract description 38
- 239000002243 precursor Substances 0.000 claims abstract description 37
- 239000013543 active substance Substances 0.000 claims abstract description 33
- 238000000034 method Methods 0.000 claims abstract description 25
- 125000003453 indazolyl group Chemical group N1N=C(C2=C1C=CC=C2)* 0.000 claims abstract description 24
- HZPSREFSUPXCMN-UHFFFAOYSA-N 1,4-dinitroimidazole Chemical compound [O-][N+](=O)C1=CN([N+]([O-])=O)C=N1 HZPSREFSUPXCMN-UHFFFAOYSA-N 0.000 claims abstract description 23
- KZNICNPSHKQLFF-UHFFFAOYSA-N succinimide Chemical compound O=C1CCC(=O)N1 KZNICNPSHKQLFF-UHFFFAOYSA-N 0.000 claims abstract description 18
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000002253 acid Substances 0.000 claims abstract description 10
- DMAYBPBPEUFIHJ-UHFFFAOYSA-N 4-bromobut-1-ene Chemical compound BrCCC=C DMAYBPBPEUFIHJ-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229960002317 succinimide Drugs 0.000 claims abstract description 8
- BAXOFTOLAUCFNW-UHFFFAOYSA-N 1H-indazole Chemical compound C1=CC=C2C=NNC2=C1 BAXOFTOLAUCFNW-UHFFFAOYSA-N 0.000 claims abstract description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 87
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 83
- 238000006243 chemical reaction Methods 0.000 claims description 67
- 238000002360 preparation method Methods 0.000 claims description 63
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-dimethylformamide Substances CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 48
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims description 24
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 18
- -1 sulfosuccinyl side chain Chemical group 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- SJRJJKPEHAURKC-UHFFFAOYSA-N N-Methylmorpholine Chemical compound CN1CCOCC1 SJRJJKPEHAURKC-UHFFFAOYSA-N 0.000 claims description 15
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 15
- VYDWQPKRHOGLPA-UHFFFAOYSA-N 5-nitroimidazole Chemical compound [O-][N+](=O)C1=CN=CN1 VYDWQPKRHOGLPA-UHFFFAOYSA-N 0.000 claims description 13
- 238000002156 mixing Methods 0.000 claims description 12
- 230000002194 synthesizing effect Effects 0.000 claims description 12
- UQZIYBXSHAGNOE-USOSMYMVSA-N Stachyose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](CO[C@@H]2[C@@H](O)[C@@H](O)[C@@H](O)[C@H](CO)O2)O1 UQZIYBXSHAGNOE-USOSMYMVSA-N 0.000 claims description 10
- UQZIYBXSHAGNOE-XNSRJBNMSA-N stachyose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO[C@@H]3[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O3)O)O2)O)O1 UQZIYBXSHAGNOE-XNSRJBNMSA-N 0.000 claims description 10
- 101100277613 Aspergillus desertorum desB gene Proteins 0.000 claims description 9
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 9
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 229960000583 acetic acid Drugs 0.000 claims description 9
- 239000012362 glacial acetic acid Substances 0.000 claims description 9
- 239000002904 solvent Substances 0.000 claims description 9
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 8
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 6
- IYCHDNQCHLMLJZ-UHFFFAOYSA-N Nicoxamat Chemical compound ONC(=O)C1=CC=CN=C1 IYCHDNQCHLMLJZ-UHFFFAOYSA-N 0.000 claims description 6
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 6
- 230000007812 deficiency Effects 0.000 claims description 6
- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 claims description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 4
- 150000007529 inorganic bases Chemical class 0.000 claims description 4
- 150000007530 organic bases Chemical class 0.000 claims description 4
- SCVFZCLFOSHCOH-UHFFFAOYSA-M potassium acetate Chemical compound [K+].CC([O-])=O SCVFZCLFOSHCOH-UHFFFAOYSA-M 0.000 claims description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 4
- GSNUFIFRDBKVIE-UHFFFAOYSA-N DMF Natural products CC1=CC=C(C)O1 GSNUFIFRDBKVIE-UHFFFAOYSA-N 0.000 claims description 3
- MHABMANUFPZXEB-UHFFFAOYSA-N O-demethyl-aloesaponarin I Natural products O=C1C2=CC=CC(O)=C2C(=O)C2=C1C=C(O)C(C(O)=O)=C2C MHABMANUFPZXEB-UHFFFAOYSA-N 0.000 claims description 3
- 239000007810 chemical reaction solvent Substances 0.000 claims description 3
- 229910000028 potassium bicarbonate Inorganic materials 0.000 claims description 3
- 235000015497 potassium bicarbonate Nutrition 0.000 claims description 3
- 239000011736 potassium bicarbonate Substances 0.000 claims description 3
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 claims description 3
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 3
- 235000017550 sodium carbonate Nutrition 0.000 claims description 3
- 235000011121 sodium hydroxide Nutrition 0.000 claims description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 2
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 claims description 2
- AHVYPIQETPWLSZ-UHFFFAOYSA-N N-methyl-pyrrolidine Natural products CN1CC=CC1 AHVYPIQETPWLSZ-UHFFFAOYSA-N 0.000 claims description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 2
- 239000012042 active reagent Substances 0.000 claims description 2
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 2
- GNOIPBMMFNIUFM-UHFFFAOYSA-N hexamethylphosphoric triamide Chemical compound CN(C)P(=O)(N(C)C)N(C)C GNOIPBMMFNIUFM-UHFFFAOYSA-N 0.000 claims description 2
- 235000011056 potassium acetate Nutrition 0.000 claims description 2
- 229940086066 potassium hydrogencarbonate Drugs 0.000 claims description 2
- 229910000160 potassium phosphate Inorganic materials 0.000 claims description 2
- 235000011009 potassium phosphates Nutrition 0.000 claims description 2
- 239000001632 sodium acetate Substances 0.000 claims description 2
- 235000017281 sodium acetate Nutrition 0.000 claims description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 2
- 239000001488 sodium phosphate Substances 0.000 claims description 2
- 229910000162 sodium phosphate Inorganic materials 0.000 claims description 2
- 235000011008 sodium phosphates Nutrition 0.000 claims description 2
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 claims description 2
- 239000003513 alkali Substances 0.000 claims 4
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 238000003786 synthesis reaction Methods 0.000 abstract description 3
- 239000000243 solution Substances 0.000 description 20
- 238000003756 stirring Methods 0.000 description 17
- 239000007864 aqueous solution Substances 0.000 description 16
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 15
- 238000001035 drying Methods 0.000 description 13
- 238000001914 filtration Methods 0.000 description 10
- 239000002244 precipitate Substances 0.000 description 10
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 8
- 239000012074 organic phase Substances 0.000 description 8
- 230000001376 precipitating effect Effects 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- GCPYCNBGGPHOBD-UHFFFAOYSA-N Delphinidin Natural products OC1=Cc2c(O)cc(O)cc2OC1=C3C=C(O)C(=O)C(=C3)O GCPYCNBGGPHOBD-UHFFFAOYSA-N 0.000 description 6
- JKHRCGUTYDNCLE-UHFFFAOYSA-O delphinidin Chemical compound [O+]=1C2=CC(O)=CC(O)=C2C=C(O)C=1C1=CC(O)=C(O)C(O)=C1 JKHRCGUTYDNCLE-UHFFFAOYSA-O 0.000 description 6
- 235000007242 delphinidin Nutrition 0.000 description 6
- 239000005457 ice water Substances 0.000 description 6
- 239000000706 filtrate Substances 0.000 description 5
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000006227 byproduct Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 229910052708 sodium Inorganic materials 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 238000010025 steaming Methods 0.000 description 4
- 238000001291 vacuum drying Methods 0.000 description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000001704 evaporation Methods 0.000 description 3
- PBGKTOXHQIOBKM-FHFVDXKLSA-N insulin (human) Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@H]1CSSC[C@H]2C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3C=CC(O)=CC=3)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3NC=NC=3)NC(=O)[C@H](CO)NC(=O)CNC1=O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O)=O)CSSC[C@@H](C(N2)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](NC(=O)CN)[C@@H](C)CC)[C@@H](C)CC)[C@@H](C)O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C=CC=CC=1)C(C)C)C1=CN=CN1 PBGKTOXHQIOBKM-FHFVDXKLSA-N 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000002390 rotary evaporation Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- DYHSDKLCOJIUFX-UHFFFAOYSA-N tert-butoxycarbonyl anhydride Chemical compound CC(C)(C)OC(=O)OC(=O)OC(C)(C)C DYHSDKLCOJIUFX-UHFFFAOYSA-N 0.000 description 3
- VUTBELPREDJDDH-UHFFFAOYSA-N 4-amino-5-hydroxymethyl-2-methylpyrimidine Chemical compound CC1=NC=C(CO)C(N)=N1 VUTBELPREDJDDH-UHFFFAOYSA-N 0.000 description 2
- 101000976075 Homo sapiens Insulin Proteins 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 125000002883 imidazolyl group Chemical group 0.000 description 2
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Substances [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- NZEDHZOVUDEBGW-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) tetradecanoate Chemical compound CCCCCCCCCCCCCC(=O)ON1C(=O)CCC1=O NZEDHZOVUDEBGW-UHFFFAOYSA-N 0.000 description 1
- ODIGIKRIUKFKHP-UHFFFAOYSA-N (n-propan-2-yloxycarbonylanilino) acetate Chemical compound CC(C)OC(=O)N(OC(C)=O)C1=CC=CC=C1 ODIGIKRIUKFKHP-UHFFFAOYSA-N 0.000 description 1
- 241000700124 Octodon degus Species 0.000 description 1
- 108010076181 Proinsulin Proteins 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- 150000001266 acyl halides Chemical class 0.000 description 1
- 230000010933 acylation Effects 0.000 description 1
- 238000005917 acylation reaction Methods 0.000 description 1
- 150000001336 alkenes Chemical group 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 150000008064 anhydrides Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 150000002473 indoazoles Chemical class 0.000 description 1
- 239000004026 insulin derivative Substances 0.000 description 1
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 235000011181 potassium carbonates Nutrition 0.000 description 1
- OVARTBFNCCXQKS-UHFFFAOYSA-N propan-2-one;hydrate Chemical compound O.CC(C)=O OVARTBFNCCXQKS-UHFFFAOYSA-N 0.000 description 1
- 238000013094 purity test Methods 0.000 description 1
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 125000000383 tetramethylene group Chemical group [H]C([H])([*:1])C([H])([H])C([H])([H])C([H])([H])[*:2] 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
- C07K14/62—Insulins
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Endocrinology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Diabetes (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
The application discloses a synthesis method of insulin, which belongs to the technical field of insulin synthesis, and particularly relates to a synthesis method of insulin, wherein tetradecanoic acid reacts with indazole or N-hydroxysulfonic acid succinimide to generate indazole side chain active substance or sulfonic acid succinyl side chain active substance, then the indazole side chain active substance or sulfonic acid succinyl side chain active substance reacts with a threo-deficiency insulin precursor desB30 under the action of functional nitroimidazole to prepare insulin, the functional nitroimidazole firstly reacts with fuming nitric acid to prepare 1, 4-dinitroimidazole, and then reacts with 4-bromo-1-butene to prepare the functional nitroimidazole; the application can also combine functional nitroimidazole with nicotinic ammoxime to prepare insulin detention. The insulin detention prepared by the method has high yield and good purity.
Description
Technical Field
The application belongs to the technical field of insulin synthesis, and particularly relates to a method for synthesizing insulin detention.
Background
Insulin detention is a novel long-acting soluble (avoiding the common side effects of insoluble insulin) insulin analogue developed by the Nor Wo Nuo g company by using genetically engineered recombinant human insulin. The structural formula is as follows: that is, an epsilon-amino group of Lys at position B29 of the human insulin analogue from which the amino acid residue at position B30 has been removed is linked to a tetradecyl fatty acid (myristic acid) side chain, wherein the B30-removed recombinant human insulin and the tetradecyl fatty acid (myristic acid) side chains are linked in the form of an amide bond.
Since proinsulin, insulin or insulin analogues all have a free alpha-amino group at amino acid position A1, B1 and a free epsilon-amino group on the side chain of lysine position B29, these three free amino groups can form covalent bonds with active esters, acyl halides or anhydrides, thereby forming mono-, di-or even tri-acylated insulin products. The acylation route disclosed in the various patents of the Norand Norde application first utilizes di-tert-butyl dicarbonate (BOC) derivative to protect the N-terminal alpha-amino groups of both chains of insulin, then utilizes myristic acid succinimidyl ester to acylate the epsilon-amino group of lysine at position B29, and finally utilizes TFA deprotection to produce insulin detes. If the acylating agent is excessive, a large amount of di-substituted or tri-substituted derivative is generated, so that the process is complicated in steps, has a large number of byproducts, increases the difficulty of downstream purification, and is unfavorable for improving the overall yield. Thus, there is a need for an improved process for synthesizing insulin detention.
Disclosure of Invention
The application aims to provide a synthesis method of insulin detention with high yield and good purity.
The technical scheme adopted by the application for achieving the purpose is as follows:
a method for synthesizing insulin detention, comprising: mixing a stachyose precursor desB30 and a side chain active substance in a reaction system, and preparing the insulin deltoid through reaction; side chain actives include indazolyl side chain actives or sulfosuccinyl side chain actives; indazolyl side chain actives have indazolyl groups and tetradecanoic acid groups, and sulfosuccinyl side chain actives have N-hydroxysulfonic acid succinimide groups and tetradecanoic acid groups.
Preferably, the reaction temperature of the reaction system is from-20 to 60 ℃; or, the reaction temperature of the reaction system is 0 to 40 ℃; or, the reaction solvent of the reaction system is at least 1 of DCM, THF, ethyl acetate, acetonitrile, DMF, DMSO, NMP, water, acetone, dimethylacetamide DMAC and hexamethylphosphoramide HMP; or the reaction solvent of the reaction system is at least 1 of DMF, DMSO, DMAC, HMP, acetonitrile-water and acetone-water.
Preferably, the reaction system comprises a base, and the base is an organic base or an inorganic base.
More preferably, the organic base comprises at least 1 of Triethylamine (TEA), diethylamine, N-methylmorpholine (NMM) and N, N-Diisopropylethylamine (DIEA); or, the inorganic base includes at least 1 of sodium acetate, potassium acetate, sodium phosphate, disodium hydrogen phosphate, potassium phosphate, dipotassium hydrogen phosphate, sodium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, and sodium hydroxide; or, the organic base includes at least 1 of Triethylamine (TEA), N-methylmorpholine (NMM) and N, N-Diisopropylethylamine (DIEA); or, the inorganic base includes at least 1 of sodium bicarbonate, sodium carbonate, potassium bicarbonate, potassium carbonate, and sodium hydroxide.
Preferably, the pH of the reaction system is from 5 to 14; or the pH value of the reaction system is 7-12.
Preferably, the reaction system contains functionalized nitroimidazole and/or nicotinic ammoxime.
More preferably, the amount of functionalized nitroimidazole used is 5-20wt% of the insulin precursor desB 30; or, the amount of the nicotinic oxime used is 1-5wt% of the insulin precursor desB 30; or the functional nitroimidazole is prepared from 4-nitroimidazole, fuming nitric acid and 4-bromo-1-butene through a series of reactions. In the process of preparing the insulin, the functional nitroimidazole can be added, the functional nitroimidazole has a nitro group, an olefin group and a cyclic structure, and under the action of the group and the structure, the synthesis of the insulin is improved, the yield of the insulin is improved, and the insulin has better purity.
Preferably, the side chain active is used in an amount of 5-20wt% of the insulin precursor desB 30.
Preferably, in the preparation of the side chain active substance, tetradecanoic acid and an active reagent are mixed in a solvent for reaction to prepare the side chain active substance; the active agent comprises indazole or N-hydroxysulfonic acid succinimide.
Preferably, in the preparation of the indazolyl side chain active, tetradecanoic acid is added into DMF, indazole is then added, EDCl is added at the temperature of 0-5 ℃, the mixture is stirred at the temperature of 20-40 ℃ for reaction for 12-24 hours, EA and water are added after the reaction is completed, an organic phase is extracted, and the organic phase is evaporated to dryness, so that the indazolyl side chain active is obtained.
More preferably, in the preparation of indazolyl side chain active, tetradecanoic acid is used in an amount of 5-20wt% of DMF.
More preferably, in the preparation of indazolyl side chain actives, indazoles are used in amounts of 50-60wt% of tetradecanoic acid.
More preferably, in the preparation of indazolyl side chain actives, EDCl is used in an amount of 80-100wt% of tetradecanoic acid.
More preferably, in the preparation of indazolyl side chain actives, EA is used in an amount of 50-150wt% of DMF.
More preferably, in the preparation of indazolyl side chain actives, water is used in an amount of 150-250wt% of DMF.
Preferably, in the preparation of the sulfosuccinyl side chain active substance, tetradecanoic acid is added into DMF, then N-hydroxysulfonic acid succinimide is added, EDCl is added at the temperature of 0-5 ℃, after mixing, stirring is carried out for 12-24 hours at the temperature of 20-40 ℃, after the reaction is completed, EA and water are added, an organic phase is extracted, and the organic phase is evaporated to dryness, so that the indazolyl side chain active substance is obtained.
More preferably, in the preparation of the sulfosuccinyl side chain active, tetradecanoic acid is used in an amount of 5 to 20wt% of DMF.
More preferably, in the preparation of the sulfosuccinyl side chain active, N-hydroxysulfonic acid succinimide is used in an amount of 80 to 120wt% of tetradecanoic acid.
More preferably, in the preparation of the sulfosuccinyl side chain active, EDCl is used in an amount of 80 to 100wt% of tetradecanoic acid.
More preferably, in the preparation of the sulfosuccinyl side chain active, EA is used in an amount of 50-150wt% of DMF.
More preferably, in the preparation of the sulfosuccinyl side chain active, water is used in an amount of 150 to 250wt% of DMF.
Preferably, in the preparation of 1, 4-dinitroimidazole, 4-nitroimidazole is mixed with glacial acetic acid, stirred for 10-30min at 10-40 ℃, fuming nitric acid and acetic anhydride are added, then stirred for 3-10min, reacted for 4-16h at 20-40 ℃, then ice water is added to separate out precipitate, and the precipitate is filtered and dried to obtain 1, 4-dinitroimidazole.
More preferably, in the preparation of 1, 4-dinitroimidazole, the amount of 4-nitroimidazole used is 10 to 40wt% of glacial acetic acid.
More preferably, in the preparation of 1, 4-dinitroimidazole, fuming nitric acid is used in an amount of 100 to 200wt% of 4-nitroimidazole.
More preferably, in the preparation of 1, 4-dinitroimidazole, acetic anhydride is used in an amount of 100 to 200wt% of 4-nitroimidazole.
More preferably, in the preparation of 1, 4-dinitroimidazole, the ice water is used in an amount of 800 to 1200wt% of glacial acetic acid.
Preferably, in the preparation of the functionalized nitroimidazole, 1, 4-dinitroimidazole is added into DMF, then 4-bromo-1-butene is added, the reaction is carried out for 48 to 96 hours at the temperature of 70 to 90 ℃, the solvent is removed by rotary evaporation after the reaction is finished, acetone is added after cooling, the mixture is kept stand for 1 to 4 hours, precipitation is carried out, and the solid is collected by filtration, thus obtaining the functionalized nitroimidazole.
More preferably, in the preparation of the functionalized nitroimidazole, the 1, 4-dinitroimidazole is used in an amount of 0.2 to 2wt% of DMF.
More preferably, in the preparation of the functionalized nitroimidazole, the amount of 4-bromo-1-butene used is from 100 to 600% by weight of 1, 4-dinitroimidazole.
More preferably, in the preparation of the functionalized nitroimidazole, the acetone is used in an amount of 40-80wt% of DMF.
Preferably, in the preparation of the insulin delsB 30, a precursor of the insulin delsB, is added into an acetonitrile aqueous solution, then functional nitroimidazole and side chain active substances are added, stirring reaction is carried out for 1-5h, after the reaction is finished, filtering is carried out, filtrate is evaporated to be nearly dry, spin-drying is carried out, a product is added into DMF, then a large amount of acetone is added, precipitation is carried out, centrifugation and vacuum drying are carried out, and the insulin delsB is obtained.
More preferably, in the preparation of insulin detention, the acetonitrile aqueous solution is formed by mixing acetonitrile and water, and the acetonitrile content of the acetonitrile aqueous solution is 40-60wt%.
More preferably, in the preparation of insulin detention, the amount of insulin precursor desB30 used is 1-4wt% of the acetonitrile aqueous solution.
More preferably, in the preparation of insulin detention, the functionalized nitroimidazole is used in an amount of 5-20wt% of the insulin precursor desB 30.
More preferably, in the preparation of insulin detention, the side chain active is used in an amount of 5-20wt% of the insulin precursor des B30.
More preferably, in the preparation of insulin detention, DMF is used in the same amount as acetonitrile in water solution, and acetone is used in an appropriate amount.
Preferably, in the preparation of the insulin delsu precursor, desB30 is added into DMF, then functional nitroimidazole and side chain active substances are added, stirring reaction is carried out for 1-5h, a large amount of acetone is added after the reaction is finished, precipitation is separated out, and the insulin delsu precursor is obtained through centrifugation and vacuum drying.
More preferably, in the preparation of insulin detention, the amount of the insulin precursor desB30 to be used is 1-4wt% of DMF.
More preferably, in the preparation of insulin detention, the functionalized nitroimidazole is used in an amount of 5-20wt% of the insulin precursor desB 30.
More preferably, in the preparation of insulin detention, the side chain active is used in an amount of 5-20wt% of the insulin precursor des B30.
More preferably, in the preparation of insulin detention, an appropriate amount of acetone is used.
In the preparation of the insulin delphinidin, the application can also add the nicotinic oxime, and the usage amount of the nicotinic oxime is 1-5wt% of the insulin delphinidin precursor desB 30. The application can combine the functional nitroimidazole and the nicotinic ammoxime when preparing the insulin, and can be used in the preparation of the insulin, thereby improving the yield of the insulin and having good purity.
The application discloses an application of functionalized nitroimidazole and/or nicotinic ammoxime in preparation of insulin, wherein the functionalized nitroimidazole contains a nitro group, an imidazole group and a butene group.
The application adopts the technical scheme that tetradecanoic acid reacts with indazole or N-hydroxysulfonic acid succinimide to generate indazole side chain active matter or sulfonic acid succinyl side chain active matter, then reacts with a threo-deficiency insulin precursor desB30 under the action of functional nitroimidazole to prepare the insulin, the functional nitroimidazole firstly reacts with fuming nitric acid to prepare 1, 4-dinitroimidazole, and then reacts with 4-bromo-1-butene to prepare the functional nitroimidazole; therefore, has the following beneficial effects: the yield of the insulin is high and the insulin has good purity. Therefore, the application is a synthesis method of the insulin digger with high yield and good purity.
Drawings
FIG. 1 is an infrared spectrogram;
FIG. 2 is a graph of the yield of insulin detention;
FIG. 3 is a chart of the purity of insulin detention.
Detailed Description
The technical scheme of the application is further described in detail below with reference to the specific embodiments and the attached drawings:
the preparation of the insulin-deficiency precursor desB30 used in the present application is referred to as follows: li Jing purification and modification process of Degu insulin and biological potency analysis [ D ], university of Zhejiang, 2019.
Example 1: synthesis method of insulin detention
Preparation of indazolyl side chain actives: tetradecanoic acid is added into DMF, indazole is then added, EDCl is added at the temperature of 0 ℃, after mixing, stirring reaction is carried out for 18 hours at the temperature of 30 ℃, EA and water are added after the reaction is finished, an organic phase is extracted, and the organic phase is evaporated to dryness, so that indazolyl side chain active substance is obtained. The amount of DMF was 100g, tetradecanoic acid was 10g, indazole was 5.69g, EDCl was 9.23g, EA was 100g, and water was 200g.
Preparation of 1, 4-dinitroimidazole: mixing 4-nitroimidazole with glacial acetic acid, stirring at 30 ℃ for 20min, adding fuming nitric acid and acetic anhydride, stirring for 5min, reacting at 30 ℃ for 8h, adding ice water to separate out precipitate, filtering, and drying to obtain 1, 4-dinitroimidazole. The amount of glacial acetic acid used was 100g, the amount of 4-nitroimidazole used was 20g, fuming nitric acid used was 30g, and acetic anhydride used was 30g. The amount of ice water used was 200g.
Preparation of functionalized nitroimidazole: adding 1, 4-dinitroimidazole into DMF, then adding 4-bromo-1-butene, reacting at 80 ℃ for 72 hours, removing solvent by rotary evaporation after the reaction is completed, cooling, adding acetone, standing for 3 hours, precipitating, filtering and collecting solids, thus obtaining the functionalized nitroimidazole. The amount of DMF used was 100g,1, 4-dinitroimidazole was 1g, 4g, and acetone was 60g.
Preparation of insulin detention: adding the delensine precursor desB30 into DMF, adding functional nitroimidazole and side chain active substances, stirring for reaction for 3 hours, adding a large amount of acetone after the reaction is finished, precipitating a precipitate, centrifuging, and drying in vacuum to obtain the insulin deltoid. The amount of DMF was 100g, the amount of desB30, the precursor of stachyose, was 2g, the amount of functional nitroimidazole was 0.2g, and the amount of side chain active was 0.2g. The proper amount of acetone is used. The side chain active is indazolyl side chain active.
Example 2: synthesis method of insulin detention
This example differs from example 1 in the preparation of insulin detention.
Preparation of insulin detention: adding the stachyose precursor desB30 into acetonitrile water solution, adding functionalized nitroimidazole and side chain active substances, stirring for reaction for 3 hours, filtering after the reaction is finished, steaming filtrate to be nearly dry, spin-drying, adding a large amount of acetone into a product, separating out precipitate, centrifuging, and drying in vacuum to obtain the insulin deltoid. The acetonitrile aqueous solution is formed by mixing acetonitrile and water, the acetonitrile content in the acetonitrile aqueous solution is 50wt%, the use amount of the acetonitrile aqueous solution is 100g, the use amount of the sodium deficiency insulin precursor desB30 is 2g, the use amount of the functional nitroimidazole is 0.2g, and the use amount of the side chain active substance is 0.2g. The DMF usage amount is the same as acetonitrile water solution, and the acetone usage amount is proper. The side chain active is indazolyl side chain active.
Example 3: synthesis method of insulin detention
Preparation of sulfosuccinyl side chain active: adding tetradecanoic acid into DMF, then adding N-hydroxysulfonic acid succinimide, adding EDCl at the temperature of 0 ℃, mixing, stirring at the temperature of 30 ℃ for reaction for 18 hours, adding EA and water after the reaction is completed, extracting an organic phase, and evaporating the organic phase to dryness to obtain indazolyl side chain active substances. The amount of DMF was 100g, the amount of tetradecanoic acid was 10g, the amount of N-hydroxysulfonic acid succinimide was 9.40g, the amount of EDCl was 9.23g, the amount of EA was 100g, and the amount of water was 200g.
Preparation of 1, 4-dinitroimidazole: mixing 4-nitroimidazole with glacial acetic acid, stirring at 30 ℃ for 20min, adding fuming nitric acid and acetic anhydride, stirring for 5min, reacting at 30 ℃ for 8h, adding ice water to separate out precipitate, filtering, and drying to obtain 1, 4-dinitroimidazole. The amount of glacial acetic acid used was 100g, the amount of 4-nitroimidazole used was 20g, fuming nitric acid used was 30g, and acetic anhydride used was 30g. The amount of ice water used was 200g.
Preparation of functionalized nitroimidazole: adding 1, 4-dinitroimidazole into DMF, then adding 4-bromo-1-butene, reacting at 80 ℃ for 72 hours, removing solvent by rotary evaporation after the reaction is completed, cooling, adding acetone, standing for 3 hours, precipitating, filtering and collecting solids, thus obtaining the functionalized nitroimidazole. The amount of DMF used was 100g,1, 4-dinitroimidazole was 1g, 4g, and acetone was 60g.
Preparation of insulin detention: adding the delensine precursor desB30 into DMF, adding functional nitroimidazole and side chain active substances, stirring for reaction for 3 hours, adding a large amount of acetone after the reaction is finished, precipitating a precipitate, centrifuging, and drying in vacuum to obtain the insulin deltoid. The amount of DMF was 100g, the amount of desB30, the precursor of stachyose, was 2g, the amount of functional nitroimidazole was 0.2g, and the amount of side chain active was 0.2g. The proper amount of acetone is used. The side chain active is sulfosuccinyl side chain active.
Example 4: synthesis method of insulin detention
This example differs from example 3 in the preparation of insulin detention.
Preparation of insulin detention: adding the stachyose precursor desB30 into acetonitrile water solution, adding functionalized nitroimidazole and side chain active substances, stirring for reaction for 3 hours, filtering after the reaction is finished, steaming filtrate to be nearly dry, spin-drying, adding a large amount of acetone into a product, separating out precipitate, centrifuging, and drying in vacuum to obtain the insulin deltoid. The acetonitrile aqueous solution is formed by mixing acetonitrile and water, the acetonitrile content in the acetonitrile aqueous solution is 50wt%, the use amount of the acetonitrile aqueous solution is 100g, the use amount of the sodium deficiency insulin precursor desB30 is 2g, the use amount of the functional nitroimidazole is 0.2g, and the use amount of the side chain active substance is 0.2g. The DMF usage amount is the same as acetonitrile water solution, and the acetone usage amount is proper. The side chain active is sulfosuccinyl side chain active.
Example 5: synthesis method of insulin detention
This example differs from example 1 in the preparation of insulin detention.
Preparation of insulin detention: adding the delensine precursor desB30 into DMF, adding functional nitroimidazole, nicotinic ammoxime and side chain active substances, stirring and reacting for 3 hours, adding a large amount of acetone after the reaction is finished, separating out precipitate, centrifuging, and drying in vacuum to obtain the insulin deltoid. The usage amount of DMF is 100g, the usage amount of the stachyose precursor desB30 is 2g, the usage amount of functional nitroimidazole is 0.2g, the usage amount of nicotinic ammoxime is 0.05g, and the usage amount of side chain active substance is 0.2g. The proper amount of acetone is used. The side chain active is indazolyl side chain active.
Example 6: synthesis method of insulin detention
This example differs from example 2 in the preparation of insulin detention.
Preparation of insulin detention: adding the stachyose precursor desB30 into acetonitrile water solution, adding functional nitroimidazole, nicotinic oxime and side chain active substances, stirring and reacting for 3 hours, filtering after the reaction is finished, steaming filtrate to be nearly dry, spin-drying, adding the product into DMF, adding a large amount of acetone, precipitating, centrifuging, and vacuum drying to obtain the insulin delphinidin. The acetonitrile aqueous solution is formed by mixing acetonitrile and water, the acetonitrile content in the acetonitrile aqueous solution is 50wt%, the use amount of the acetonitrile aqueous solution is 100g, the use amount of the sodium deficiency insulin precursor desB30 is 2g, the use amount of the functional nitroimidazole is 0.2g, the use amount of the nicotinic ammoxime is 0.05g, and the use amount of the side chain active substance is 0.2g. The DMF usage amount is the same as acetonitrile water solution, and the acetone usage amount is proper. The side chain active is indazolyl side chain active.
Example 7: synthesis method of insulin detention
This example differs from example 3 in the preparation of insulin detention.
Preparation of insulin detention: adding the delensine precursor desB30 into DMF, adding functional nitroimidazole, nicotinic ammoxime and side chain active substances, stirring and reacting for 3 hours, adding a large amount of acetone after the reaction is finished, separating out precipitate, centrifuging, and drying in vacuum to obtain the insulin deltoid. The usage amount of DMF is 100g, the usage amount of the stachyose precursor desB30 is 2g, the usage amount of functional nitroimidazole is 0.2g, the usage amount of nicotinic ammoxime is 0.05g, and the usage amount of side chain active substance is 0.2g. The proper amount of acetone is used. The side chain active is sulfosuccinyl side chain active.
Example 8: synthesis method of insulin detention
This example differs from example 4 in the preparation of insulin detention.
Preparation of insulin detention: adding the stachyose precursor desB30 into acetonitrile water solution, adding functional nitroimidazole, nicotinic oxime and side chain active substances, stirring and reacting for 3 hours, filtering after the reaction is finished, steaming filtrate to be nearly dry, spin-drying, adding the product into DMF, adding a large amount of acetone, precipitating, centrifuging, and vacuum drying to obtain the insulin delphinidin. The acetonitrile aqueous solution is formed by mixing acetonitrile and water, the acetonitrile content in the acetonitrile aqueous solution is 50wt%, the use amount of the acetonitrile aqueous solution is 100g, the use amount of the sodium deficiency insulin precursor desB30 is 2g, the use amount of the functional nitroimidazole is 0.2g, the use amount of the nicotinic ammoxime is 0.05g, and the use amount of the side chain active substance is 0.2g. The DMF usage amount is the same as acetonitrile water solution, and the acetone usage amount is proper. The side chain active is sulfosuccinyl side chain active.
Test example:
the application has carried out infrared characterization on the functionalized nitroimidazole prepared in the example 1, the result is shown in figure 1, wherein, the temperature is 3147cm -1 At 2882cm is the infrared absorption peak of=c-H on the imidazole ring -1 The peak of the infrared absorption of methylene was at 1572cm -1 、1478cm -1 Infrared absorption peaks c=c and c=n, 1318cm -1 Is C-NO 2 Is 1273cm -1 Is N-NO 2 The infrared absorption peak of (2) indicates that the functionalized nitroimidazole is obtained.
The application calculates the yield of the insulin delphinidin prepared by the method of examples 1-8, wherein the result is shown in figure 2, S1 is example 1, S2 is example 2, S3 is example 3, S4 is example 4, S5 is example 5, S6 is example 6, S7 is example 7, and S8 is example 8; the application discovers that the functionalized nitroimidazole can be applied to the preparation of the insulin detention, and the functionalized nitroimidazole can not be removed by evaporation, but in the mixture containing the functionalized nitroimidazole and the insulin detention, the functionalized nitroimidazole can be dissolved in a solvent, and then a large amount of solvent capable of precipitating the insulin detention is added, so that the insulin detention is precipitated, the insulin detention is further separated, and the high-yield insulin detention product can be obtained by applying the functionalized nitroimidazole in the method; the reaction system of the insulin digger comprises a reaction system using acetonitrile water solution and a reaction system using DMF solution, and the reaction system using acetonitrile water solution is superior to the reaction system using DMF solution, which shows that the functionalized nitroimidazole has better effect when being applied to the reaction system using acetonitrile water solution; in the reaction system for preparing the insulin, the functionalized nitroimidazole and the nicotinic ammoxime can be used together, and the yield of the insulin is higher after the functionalized nitroimidazole and the nicotinic ammoxime are added into the preparation of the insulin.
The purity test is carried out on the insulin delphinidin prepared by the method of the examples 1-8, the result is shown in figure 3, wherein S1 is the example 1, S2 is the example 2, S3 is the example 3, S4 is the example 4, S5 is the example 5, S6 is the example 6, S7 is the example 7, and S8 is the example 8; the application discovers that the functionalized nitroimidazole can be applied to the preparation of the insulin detention, and the functionalized nitroimidazole can not be removed by evaporation, but in the mixture containing the functionalized nitroimidazole and the insulin detention, the functionalized nitroimidazole can be dissolved in a solvent, and then a large amount of solvent capable of precipitating the insulin detention is added, so that the insulin detention is precipitated, the insulin detention is further separated, and the high-purity insulin detention product can be obtained by applying the functionalized nitroimidazole in the method; the reaction system of the insulin digger comprises a reaction system using acetonitrile water solution and a reaction system using DMF solution, and the reaction system using acetonitrile water solution is superior to the reaction system using DMF solution, which shows that the functionalized nitroimidazole has better effect when being applied to the reaction system using acetonitrile water solution; in the reaction system for preparing the insulin detention, the functionalized nitroimidazole and the nicotinic ammoxime can be used together, and the purity of the obtained insulin detention is higher after the functionalized nitroimidazole and the nicotinic ammoxime are added into the preparation of the insulin detention. The higher purity of the insulin niter indicates that the byproduct generation rate is low, and the functional nitroimidazole used in the application can reduce the byproduct generation, and the functional nitroimidazole and the nicotinic ammoxime are used together, so that the byproduct generation can be further reduced.
The above embodiments are merely for illustrating the present application and not for limiting the same, and various changes and modifications may be made by one of ordinary skill in the art without departing from the spirit and scope of the application. Therefore, all equivalent technical solutions are also within the scope of the present application, which is defined by the claims.
Claims (9)
1. A method for synthesizing insulin detention, comprising: mixing a stachyose precursor desB30 and a side chain active substance in a reaction system, and preparing the insulin deltoid through reaction; the side chain actives include indazolyl side chain actives or sulfosuccinyl side chain actives; indazolyl side chain actives have indazolyl groups and tetradecanoic acid groups, and sulfosuccinyl side chain actives have N-hydroxysulfonic acid succinimide groups and tetradecanoic acid groups; in the preparation of the functional nitroimidazole, 4-nitroimidazole and fuming nitric acid react in glacial acetic acid containing acetic anhydride to prepare 1, 4-dinitroimidazole, and then 1, 4-dinitroimidazole and 4-bromo-1-butene react in DMF to prepare the functional nitroimidazole;
the usage amount of 4-nitroimidazole is 10-40wt% of glacial acetic acid, the usage amount of fuming nitric acid is 100-200wt% of 4-nitroimidazole, and the usage amount of acetic anhydride is 100-200wt% of 4-nitroimidazole;
the use amount of the 1, 4-dinitroimidazole is 0.2-2wt% of DMF, and the use amount of the 4-bromo-1-butene is 100-600wt% of the 1, 4-dinitroimidazole.
2. The method for synthesizing insulin detention according to claim 1, wherein the method comprises the following steps: the reaction temperature of the reaction system is-20 to 60 ℃; or the pH value of the reaction system is 5-14; or, the reaction solvent of the reaction system is at least 1 of DCM, THF, ethyl acetate, acetonitrile, DMF, DMSO, NMP, water, acetone, dimethylacetamide DMAC and hexamethylphosphoramide HMP.
3. The method for synthesizing insulin detention according to claim 1, wherein the method comprises the following steps: the reaction system comprises alkali, wherein the alkali is organic alkali or inorganic alkali.
4. A method of synthesizing insulin detention according to claim 3, wherein: the organic base comprises at least 1 of triethylamine, diethylamine, N-methylmorpholine and N, N-diisopropylethylamine; or, the inorganic base includes at least 1 of sodium acetate, potassium acetate, sodium phosphate, disodium hydrogen phosphate, potassium phosphate, dipotassium hydrogen phosphate, sodium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, and sodium hydroxide.
5. The method for synthesizing insulin detention according to claim 2, wherein: the reaction temperature of the reaction system is 0 to 40 ℃; or the pH value of the reaction system is 7-12.
6. The method for synthesizing insulin detention according to claim 1, wherein the method comprises the following steps: the reaction system contains nicotinic ammoxime.
7. The method for synthesizing insulin detention according to claim 6, wherein: the usage amount of the functional nitroimidazole is 5-20wt% of the insulin deficiency precursor desB 30; alternatively, the amount of nicotinic oxime used is 1-5wt% of the insulin precursor desB 30.
8. The method for synthesizing insulin detention according to claim 1, wherein the method comprises the following steps: the side chain active substance is used in an amount of 5-20wt% of the insulin deficiency precursor desB 30.
9. The method for synthesizing insulin detention according to claim 1, wherein the method comprises the following steps: in the preparation of the side chain active substance, tetradecanoic acid and an active reagent are mixed in a solvent for reaction to prepare the side chain active substance; the active agent comprises an indazole or an N-hydroxysulfonic acid succinimide.
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