JPH0224836B2 - - Google Patents
Info
- Publication number
- JPH0224836B2 JPH0224836B2 JP1086538A JP8653889A JPH0224836B2 JP H0224836 B2 JPH0224836 B2 JP H0224836B2 JP 1086538 A JP1086538 A JP 1086538A JP 8653889 A JP8653889 A JP 8653889A JP H0224836 B2 JPH0224836 B2 JP H0224836B2
- Authority
- JP
- Japan
- Prior art keywords
- added
- solvent
- pro
- dmf
- mtr
- 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.)
- Expired - Lifetime
Links
- 238000004519 manufacturing process Methods 0.000 claims description 39
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 26
- -1 4-methoxy-2,3,6-trimethylbenzenesulfonyl group Chemical group 0.000 claims description 25
- 125000006239 protecting group Chemical group 0.000 claims description 22
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 13
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 11
- 238000009833 condensation Methods 0.000 claims description 7
- 230000005494 condensation Effects 0.000 claims description 7
- 125000002883 imidazolyl group Chemical group 0.000 claims description 7
- QMMFVYPAHWMCMS-UHFFFAOYSA-N Dimethyl sulfide Chemical compound CSC QMMFVYPAHWMCMS-UHFFFAOYSA-N 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 6
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 claims description 3
- 230000008030 elimination Effects 0.000 claims description 3
- 238000003379 elimination reaction Methods 0.000 claims description 3
- HNKJADCVZUBCPG-UHFFFAOYSA-N thioanisole Chemical compound CSC1=CC=CC=C1 HNKJADCVZUBCPG-UHFFFAOYSA-N 0.000 claims description 2
- 230000001012 protector Effects 0.000 claims 1
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 114
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 96
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 93
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 77
- 239000002904 solvent Substances 0.000 description 77
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 56
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 54
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 53
- 238000001914 filtration Methods 0.000 description 44
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 36
- 239000000203 mixture Substances 0.000 description 36
- 238000000921 elemental analysis Methods 0.000 description 35
- 238000002844 melting Methods 0.000 description 34
- 230000008018 melting Effects 0.000 description 34
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 30
- 239000000843 powder Substances 0.000 description 28
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 27
- AYMLQYFMYHISQO-QMMMGPOBSA-N (2s)-3-(1h-imidazol-3-ium-5-yl)-2-[(2-methylpropan-2-yl)oxycarbonylamino]propanoate Chemical compound CC(C)(C)OC(=O)N[C@H](C(O)=O)CC1=CN=CN1 AYMLQYFMYHISQO-QMMMGPOBSA-N 0.000 description 26
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 26
- 238000010531 catalytic reduction reaction Methods 0.000 description 23
- 239000013078 crystal Substances 0.000 description 21
- XBPCUCUWBYBCDP-UHFFFAOYSA-N Dicyclohexylamine Chemical compound C1CCCCC1NC1CCCCC1 XBPCUCUWBYBCDP-UHFFFAOYSA-N 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 17
- 238000000034 method Methods 0.000 description 16
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 15
- 235000017557 sodium bicarbonate Nutrition 0.000 description 15
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 13
- ZUSSTQCWRDLYJA-UHFFFAOYSA-N n-hydroxy-5-norbornene-2,3-dicarboximide Chemical compound C1=CC2CC1C1C2C(=O)N(O)C1=O ZUSSTQCWRDLYJA-UHFFFAOYSA-N 0.000 description 13
- 239000003208 petroleum Substances 0.000 description 13
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 11
- 238000003756 stirring Methods 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- YEDUAINPPJYDJZ-UHFFFAOYSA-N 2-hydroxybenzothiazole Chemical compound C1=CC=C2SC(O)=NC2=C1 YEDUAINPPJYDJZ-UHFFFAOYSA-N 0.000 description 8
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 8
- 238000010306 acid treatment Methods 0.000 description 8
- 238000001816 cooling Methods 0.000 description 7
- 239000002244 precipitate Substances 0.000 description 7
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N hydrochloric acid Substances Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 6
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 5
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 description 5
- PUBCCFNQJQKCNC-XKNFJVFFSA-N gastrin-releasingpeptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(N)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)CNC(=O)[C@H](C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)C(C)C)[C@@H](C)O)C(C)C)[C@@H](C)O)C(C)C)C1=CNC=N1 PUBCCFNQJQKCNC-XKNFJVFFSA-N 0.000 description 5
- OOFAEFCMEHZNGP-UHFFFAOYSA-N 1-n',1-n'-dimethylpropane-1,1-diamine Chemical compound CCC(N)N(C)C OOFAEFCMEHZNGP-UHFFFAOYSA-N 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- PAFZNILMFXTMIY-UHFFFAOYSA-N cyclohexylamine Chemical compound NC1CCCCC1 PAFZNILMFXTMIY-UHFFFAOYSA-N 0.000 description 4
- 150000002148 esters Chemical class 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 229940024606 amino acid Drugs 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 3
- 229940098779 methanesulfonic acid Drugs 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- AWONIZVBKXHWJP-UHFFFAOYSA-N 1-methoxy-2,3,5-trimethylbenzene Chemical compound COC1=CC(C)=CC(C)=C1C AWONIZVBKXHWJP-UHFFFAOYSA-N 0.000 description 2
- OGRAOKJKVGDSFR-UHFFFAOYSA-N 2,3,5-trimethylphenol Chemical compound CC1=CC(C)=C(C)C(O)=C1 OGRAOKJKVGDSFR-UHFFFAOYSA-N 0.000 description 2
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- 108090001053 Gastrin releasing peptide Proteins 0.000 description 2
- 102100036519 Gastrin-releasing peptide Human genes 0.000 description 2
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 2
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- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 2
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 2
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- 238000000354 decomposition reaction Methods 0.000 description 2
- MDKXBBPLEGPIRI-UHFFFAOYSA-N ethoxyethane;methanol Chemical compound OC.CCOCC MDKXBBPLEGPIRI-UHFFFAOYSA-N 0.000 description 2
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- WWECJGLXBSQKRF-UHFFFAOYSA-N n,n-dimethylformamide;methanol Chemical compound OC.CN(C)C=O WWECJGLXBSQKRF-UHFFFAOYSA-N 0.000 description 2
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- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 2
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- 108010071635 tyrosyl-prolyl-arginine Proteins 0.000 description 2
- JXGVXCZADZNAMJ-NSHDSACASA-N (2s)-1-phenylmethoxycarbonylpyrrolidine-2-carboxylic acid Chemical compound OC(=O)[C@@H]1CCCN1C(=O)OCC1=CC=CC=C1 JXGVXCZADZNAMJ-NSHDSACASA-N 0.000 description 1
- WCOJOHPAKJFUDF-LBPRGKRZSA-N (2s)-3-(1h-imidazol-5-yl)-2-(phenylmethoxycarbonylamino)propanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC=1C=CC=CC=1)C1=CN=CN1 WCOJOHPAKJFUDF-LBPRGKRZSA-N 0.000 description 1
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- 238000007796 conventional method Methods 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 description 1
- WEVKQMBWAPJFSP-UHFFFAOYSA-N ethyl acetate;2-pyridin-2-ylacetic acid;hydrate Chemical compound O.CCOC(C)=O.OC(=O)CC1=CC=CC=N1 WEVKQMBWAPJFSP-UHFFFAOYSA-N 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 150000004820 halides Chemical class 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- BXWNKGSJHAJOGX-UHFFFAOYSA-N hexadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCO BXWNKGSJHAJOGX-UHFFFAOYSA-N 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 150000002410 histidine derivatives Chemical class 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 229910000040 hydrogen fluoride Inorganic materials 0.000 description 1
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- UZKWTJUDCOPSNM-UHFFFAOYSA-N methoxybenzene Substances CCCCOC=C UZKWTJUDCOPSNM-UHFFFAOYSA-N 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- WHQSYGRFZMUQGQ-UHFFFAOYSA-N n,n-dimethylformamide;hydrate Chemical compound O.CN(C)C=O WHQSYGRFZMUQGQ-UHFFFAOYSA-N 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 108010004914 prolylarginine Proteins 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- UQDJGEHQDNVPGU-UHFFFAOYSA-N serine phosphoethanolamine Chemical compound [NH3+]CCOP([O-])(=O)OCC([NH3+])C([O-])=O UQDJGEHQDNVPGU-UHFFFAOYSA-N 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000012312 sodium hydride Substances 0.000 description 1
- 229910000104 sodium hydride Inorganic materials 0.000 description 1
- XHFLOLLMZOTPSM-UHFFFAOYSA-M sodium;hydrogen carbonate;hydrate Chemical compound [OH-].[Na+].OC(O)=O XHFLOLLMZOTPSM-UHFFFAOYSA-M 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
- 101150066801 tea1 gene Proteins 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- ITMCEJHCFYSIIV-UHFFFAOYSA-N triflic acid Chemical compound OS(=O)(=O)C(F)(F)F ITMCEJHCFYSIIV-UHFFFAOYSA-N 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Description
本発明は、ヒスチジンのイミダゾール基を保護
することによるペプチドの製造法またはその塩に
関する。
ヒスチジンを含むペプチドを合成するに当つて
は、イミダゾール基が中性付近で解離して弱い塩
基性を示すため、種々の技術的に困難な問題、た
とえば、縮合時のイミダゾール基へのアシル化、
縮合後の精製の困難さ、ラセミ化のしやすさなど
を起こすことが多く、その有用な保護基が望まれ
ていた。
該保護基としては、α−アミノ保護基と選択的
に切断できることが望ましく、トリフルオロ酢酸
処理、あるいは接触還元に安定であり、かつ緩和
な方法で除去できるものが望ましい。
従来からの保護基としては、ベンジル基、カル
ボベンゾキシ基等が知られているが、これらは接
触還元で除去されるため、従つてα−アミノ保護
基としてはカルボベンゾキシ基を用いることがで
きない欠点がある。接触還元、トリフルオロ酢酸
処理の両者に安定な保護基としては、従来よりパ
ラトルエンスルホニル基が知られているが、この
保護基は、意外にも塩基性条件では非常に不安定
で、酸処理したあとの中和操作時（たとえばトリ
エチルアミンで中和する時）に、かなりの割合で
その保護基の脱離が認められることが多い。この
種の保護基を改良した保護基としては、４−メト
キシベンゼンスルホニル基が、最近知られている
が、この保護基の場合においても、塩基性条件下
における安定性は不十分である。本発明者らは
種々の置換ベンゼンスルホニル型の保護基を検討
した結果、４−メトキシ−２，３，６−トリメチ
ルベンゼンスルホニル基が、接触還元、トリフル
オロ酢酸処理、トリエチルアミン等の塩基性条件
下で十分安定であり、かつ、緩和な酸処理によつ
て、除去できることを見い出し、さらに研究した
結果、本発明を完成した。
すなわち、本発明は、ヒスチジンを有するペプ
チドの製造にあたり、ヒスチジンのイミダゾール
基を、４−メトキシ−２，３，６−トリメチルベ
ンゼンスルホニル基で保護し、ペプチド縮合した
のち、該保護基を酸または１−ヒドロキシベンゾ
トリアゾールで脱離せしめることを特徴とするペ
プチドの製造法である。
本発明においては、一般式
〔式中、Ｒは水素またはα−アミノ基の保護基
を、４−メトキシ−２，３，６−トリメチルベン
ゼンスルホニル基は（Ａ）環の１または３位のい
ずれかのＮに結合していることを表わす〕で示さ
れるヒスチジン誘導体およびその塩が用いられ
る。
本発明で用いられる４−メトキシ−２，３，６
−トリメチルベンゼンスルホニル基は新規な保護
基であり、通常、ハロゲニドの形で使用に供せら
れ、たとえば参考例１の方法で製造される。
本発明において、４−メトキシ−２，３，６−
トリメチルベンゼンスルホニル基で保護されたイ
ミダゾール基を有するヒスチジンは、たとえば次
の常套手段で製造できる。すなわち、４−メトキ
シ−２，３，６−トリメチルベンゼンスルホニル
ハライド、好ましくはクロライドと、α−アミノ
基を保護したヒスチジンを、反応させることによ
り製造できる。この反応は、たとえば、約−10°
〜＋50℃の範囲の適宜の温度で、溶媒（たとえ
ば、水、含水テトラヒドロフラン、含水ジオキサ
ン、含水アセトン、含水アセトニトリル、含水ジ
メチルホルムアミド）を使用して実施してもよ
い。塩基としては、種々の塩基を用いることがで
きるが、その中でも、炭酸水素ナトリウム、炭酸
ナトリウム、トリエチルアミン等が好ましい。α
−アミノ基の保護基としては、公知の保護基、た
とえば、カルボベンゾキシ基、ｐ−ニトロベンジ
ルオキシカルボニル基、ｐ−メトキシベンジルオ
キシカルボニル基、ｔ−ブトキシカルボニル基、
ｔ−アミロキシカルボニル基、９−フルオレニル
メトキシカルボニル基、イソニコチニルオキシカ
ルボニル基、ｏ−ニトロフエニルスルフエニル
基、２−（ｐ−ビフエニル）イソプロピルオキシ
カルボニル基などが使用でき、そのなかでも、ヒ
スチジンのα−アミノ基をカルボベンゾキシ基ま
たは、ｔ−ブトキシカルボニル基で保護したもの
が有利に用いられ、ジシクロヘキシルアミン、シ
クロヘキシルアミン、ナトリウムなどの塩として
ペプチド縮合に供せられる。
このようにして得られる４−メトキシ−２，
３，６−トリメチルベンゼンスルホニル基で保護
されたイミダゾール基を有するヒスチジンは、該
アミノ酸残基を有するペプチド縮合反応に常套手
段により、極めて有利に使用しうる。この常套手
段としては、例えばM.Bodansky及びM.A.
Ondetti著、ペプチド・シンセシス（Peptide
Synthesis）、Inter science，New York，1966
年；F.M.Finn及びK.Hofmann著 ザ・プロテイ
ンズ（The Proteins）、第２巻、H.Neurath，R.
L.Hiee編集、Academic Press Inc.New York，
1976年；泉屋信夫他著“ペプチド合成”丸善(株)
1975年などに記載された方法、たとえばアジド
法、クロライド法、酸無水物法、混酸無水物法、
DCC法、活性エステル法、ウツドワード試薬Ｋ
を用いる方法、カルボジイミダゾール法、酸化還
元法、DCC／HONB法などが挙げられる。
ところで、ペプチド縮合をするためには、α−
アミノ保護基がｔ−ブトキシカルボニル基の場合
にはトリフルオロ酢酸等で酸処理したのち、トリ
エチルアミン等の塩基による中和が必要である
が、従来、良く用いられているパラトルエンスル
ホニル基は、上記の塩基性条件下では、極めて不
安定であり、又、パラメトキシベンゼンスルホニ
ル基においても、その安定性は不十分である。こ
れに比べて、４−メトキシ−２，３，６−トリメ
チルベンゼンスルホニル基は、後述の実施例にも
示す通り極めて安定であり、その有用性は極めて
高い。
次に、ペプチド縮合後、本発明の保護基を、酸
または１−ヒドロキシベンゾトリアゾールによつ
て除去する。
酸による脱離方法としては、無水弗化水素、メ
タンスルホン酸、トリフルオロメタンスルホン酸
等の公知の酸処理方法が適用できる。さらに、本
発明方法の場合には、新しい酸処理方法として、
少量のメタンスルホン酸を含有するトリフルオロ
酢酸またはトリフルオロ酢酸が有利に使用でき、
特にチオアニソール、アニソールまたはジメチル
スルフイドの存在下で行なうと、脱離作用が有利
に進行する。
以上に述べたごとく、本発明の保護基は、接触
還元、トリフルオロ酢酸等の酸に対して安定であ
り、かつ、トリエチルアミン等の塩基に対しても
安定であるなど、従来のイミダゾール基の保護基
には認められない特徴を有している。
次に、本発明を参考例および実施例をあげて、
さらに詳しく説明する。なお、本明細書において
はアミノ酸、ペプチド、保護基、活性基等に関
し、IUPAC−IUB commission on Biological
Nomenclatureに基づく略号あるいは当該分野に
おける慣用略号で表示する場合がある。それらを
以下に例示する。Trp：トリプトフアン；Lys：
リジン；His：ヒスチジン；Arg：アルギニン；
Ser：セリン；Gly：グリシン；Ala：アラニン；
Pro：プロリン；Thr：スレオニン；Gln：グル
タミン；Val：バリン；Leu：ロイシン；Ile：イ
ソロイシン；Met：メチオニン；Tyr：チロシン
（以上、特に表示のない場合はアミノ酸はＬ体を
さすものとする。但しGlyを除く）；Ｚ：カルボ
ベンゾキシ；Boc：ｔ−ブトキシカルボニル；
OBut：ｔ−ブチルエステル；HONBおよび
ONB：Ｎ−ハイドロキシ−５−ノルボルネン−
２，３−ジカルボキシイミドおよびそのエステ
ル；HOBt：Ｎ−ハイドロキシベンヅトリアゾー
ル；DCC：Ｎ，N′−ジシクロヘキシルカルボジ
イミド；DCU：Ｎ，N′−ジシクロヘキシルウレ
ア；Mbs：４−メトキシベンゼンスルホニル：
Pme：ペンタメチルベンゼンスルホニル；Mtr：
４−メトキシ−２，３，６−トリメチルベンゼン
スルホニル；CHA：シクロヘキシルアミン；
DCHA：ジシクロヘキシルアミン；DMF：ジメ
チルホルムアミド；TEA：トリエチルアミン；
また本明細書に示す薄層クロマトグラフイーの展
開溶媒は、Rf1：クロロホルム−メタノール−酢
酸（９：１：0.5）；Rf2：酢酸エチル−ピリジン
−酢酸−水（60：20：６：10）；Rf3：クロロホ
ルム−メタノール−水（７：３：0.5）；Rf4：ｎ
−ブタノール−ピリジン−酢酸−水（30：20：
６：24）である。
参考例 １
(1) ２，３，５−トリメチルアニソールの合成
２，３，５−トリメチルフエノール10ｇ、沃化
メチル10.4mlをジメチルスルフオキシド100mlに
とかし、氷冷し、これに、60％油性水素化ナトリ
ウム5.6ｇを加え、10時間かきまぜた。これに水
を加えたのち、エーテルで抽出し、エーテル層は
水洗し、無水硫酸ナトリウムで乾燥した。溶媒を
留去して、油状物を得た。収量12.9ｇ（定量的）。
(2) ４−メトキシ−２，３，６−トリメチルベン
ゼンスルフオニルクロリドの合成
２，３，５−トリメチルアニソール4.5ｇを、
塩化メチレン500mlにとかし、−5゜〜−10℃に冷却
したのち、クロルスルフオン酸6.0mlを含む塩化
メチレン溶液400mlを滴下した。その後、室温に
までもどし、５％炭酸水素ナトリウム水を含む氷
上にあけた。塩化メチレン層は水洗したのち、無
水硫酸マグネシウムで乾燥した。溶媒を留去した
のち、ｎ−ヘキサンより結晶として、ろ取した。
収量5.0ｇ（67.0％）
融点 56−58℃
元素分析 C10H13O3SClとして
計算値：Ｃ 48.29；Ｈ 5.27；Ｓ 12.89
；Cl 14.26
実験値：Ｃ 48.42；Ｈ 5.21；Ｓ 12.61
；Cl 14.25
参考例 ２
BOC−His（Mbs）−OH・DCHAの製造
BOC−His−OH1.50ｇを水10ml、アセトン10
mlの混合液にとかし、氷冷した。これに
TEA1.65mlを加えたのち、激しくかきまぜなが
ら、Mbs−Cl1.21ｇをアセトンにとかした液10ml
を滴下し、室温で１時間撹拌した。クエン酸を加
えたのち、アセトンを留去し、酢酸エチルで抽出
し、無水硫酸ナトリウムを用いて乾燥した。
DCHA 1.06mlを加えて、溶媒を留去したのち、
残留油状物を冷蔵庫内で一夜放置すると結晶化す
るので、エーテルを加えて、これをろ取した。収
量 2.65ｇ（74.0％） 融点 146−147℃
〔α〕23 D＋21.0゜（Ｃ＝0.93、メタノール）
元素分析 C30H46O7N4Sとして
計算値：Ｃ 59.38；Ｈ 7.64；Ｎ 9.23
；Ｓ 5.29
実験値：Ｃ 59.16；Ｈ 7.70；Ｎ 8.95
；Ｓ 5.28
参考例 ３
BOC−His（Mtr）−OH・DCHAの製造
BOC−His−OH5.11ｇを水30ml、アセトン30
mlの混合液にとかし、氷冷した。これに
TEA5.60ml１を加えたのち、激しくかきまぜな
がら、Mtr−Cl4.97ｇをアセトンにとかした液30
mlを滴下し、そのまゝ２時間かきまぜた。アセト
ンを留去したのちクエン酸酸性として、酢酸エチ
ルで抽出し、無水硫酸ナトリウムを用いて乾燥し
た。DCHA3.60mlを加えたのち、溶媒を留去し、
残留油状物を冷蔵庫内で一夜放置すると、結晶化
するのでエーテルを加えて再結晶として、これを
ろ取した。収量7.80ｇ（60.1％）融点 136−137
℃
〔α〕23 D＋18.8゜（Ｃ＝1.01、メタノール）、Rf10.63
元素分析 C21H29O7N3S・C12H23Nとして
計算値：Ｃ 61.08；Ｈ 8.08；Ｎ 8.64
；Ｓ 4.94
実験値：Ｃ 61.19；Ｈ 8.05；Ｎ 8.89
；Ｓ 4.73
参考例 ４
Ｚ−His（Mtr）−OH・DCHAの製造
Ｚ−His−OH2.90ｇを水15ml、アセトン15ml
の混合液にとかし氷冷した。これにTEA2.80ml
を加えたのち激しくかきまぜながら、Mtr−
Cl2.49ｇをアセトンにとかした液15mlを滴下し、
そのまゝ約１時間かきまぜた。アセトンを留去し
たのち、クエン酸酸性として、酢酸エチルで抽出
し、無水硫酸ナトリウムを用いて乾燥した。
DCHA2mlを加えたのち、溶媒を留去し、エーテ
ルを加えて結晶後これをろ取した。
収量 4.10ｇ（60.0％）、融点149−150℃
〔α〕24 D＋14.7゜（Ｃ＝0.87、メタノール）、
Rf1 0.62
元素分析 C24H27O7N3S・C12H23Nとして
計算値：Ｃ 63.32；Ｈ 7.38；Ｎ 8.21
；Ｓ 4.07
実験値：Ｃ 62.94；Ｈ 7.11；Ｎ 8.11
；Ｓ 4.80
試験例 ４−メトキシ−２，３，６−トリメチル
ベンゼンスルホニル基（Mtr）と４−メトキシベ
ンゼンスルホニル基（Mbs）、パラトルエンスル
ホニル基（Tos）の塩基性条件下での安定性の比
較
BOC−His（Mtr）−OH・DCHA，BOC−His
（Mbs）−OH・DCHA，BOC−His（Tos）−
OH・DCHAを各100mgを取り、0.5N−トリエチ
ルアミンを含む50％含水メタノールにとかし、26
℃で一定時間放置したのち、高速液体クロマトグ
ラフイーを用いて残存する原料を定量した。別表
に示すごとく、Mtr基は他の保護基（Mbs，
Tos）に比較して、はるかに安定であり、その有
用性を確認することができた。
The present invention relates to a method for producing a peptide or a salt thereof by protecting the imidazole group of histidine. When synthesizing peptides containing histidine, the imidazole group dissociates near neutrality and exhibits weak basicity, so there are various technically difficult problems such as acylation of the imidazole group during condensation,
Since they often cause difficulties in purification after condensation and are easily racemized, a useful protecting group has been desired. The protecting group is preferably one that can selectively cleave the α-amino protecting group, is stable to trifluoroacetic acid treatment or catalytic reduction, and can be removed by a mild method. As conventional protecting groups, benzyl group, carbobenzoxy group, etc. are known, but since these are removed by catalytic reduction, it is therefore not possible to use carbobenzoxy group as the α-amino protecting group. There is a drawback that it cannot be done. Para-toluenesulfonyl group has been known as a protecting group that is stable against both catalytic reduction and trifluoroacetic acid treatment, but surprisingly this protecting group is very unstable under basic conditions and is not susceptible to acid treatment. During the subsequent neutralization operation (for example, when neutralizing with triethylamine), a considerable amount of the protecting group is often removed. A 4-methoxybenzenesulfonyl group has recently been known as an improved protecting group of this type, but even in the case of this protecting group, stability under basic conditions is insufficient. The present inventors investigated various substituted benzenesulfonyl-type protecting groups and found that the 4-methoxy-2,3,6-trimethylbenzenesulfonyl group was protected under basic conditions such as catalytic reduction, trifluoroacetic acid treatment, and triethylamine. They found that it is sufficiently stable and can be removed by mild acid treatment, and as a result of further research, they completed the present invention. That is, in producing a peptide having histidine, the present invention protects the imidazole group of histidine with a 4-methoxy-2,3,6-trimethylbenzenesulfonyl group, performs peptide condensation, and then converts the protecting group into an acid or - A method for producing a peptide, which is characterized by elimination with hydroxybenzotriazole. In the present invention, the general formula [In the formula, R is hydrogen or a protecting group for the α-amino group, and the 4-methoxy-2,3,6-trimethylbenzenesulfonyl group is bonded to N at either the 1st or 3rd position of the (A) ring. Histidine derivatives and salts thereof are used. 4-methoxy-2,3,6 used in the present invention
The -trimethylbenzenesulfonyl group is a novel protecting group, which is usually used in the form of a halide, and is produced, for example, by the method of Reference Example 1. In the present invention, 4-methoxy-2,3,6-
Histidine having an imidazole group protected with a trimethylbenzenesulfonyl group can be produced, for example, by the following conventional method. That is, it can be produced by reacting 4-methoxy-2,3,6-trimethylbenzenesulfonyl halide, preferably chloride, with histidine with an α-amino group protected. This reaction is, for example, approximately −10°
It may be carried out using a solvent (eg, water, aqueous tetrahydrofuran, aqueous dioxane, aqueous acetone, aqueous acetonitrile, aqueous dimethylformamide) at a suitable temperature in the range of ~+50°C. Various bases can be used as the base, and among them, sodium hydrogen carbonate, sodium carbonate, triethylamine, etc. are preferred. α
- As the protecting group for the amino group, known protecting groups such as carbobenzoxy group, p-nitrobenzyloxycarbonyl group, p-methoxybenzyloxycarbonyl group, t-butoxycarbonyl group,
t-amyloxycarbonyl group, 9-fluorenylmethoxycarbonyl group, isonicotinyloxycarbonyl group, o-nitrophenylsulfenyl group, 2-(p-biphenyl)isopropyloxycarbonyl group, etc. can be used, among which However, histidine in which the α-amino group is protected with a carbobenzoxy group or a t-butoxycarbonyl group is advantageously used, and is subjected to peptide condensation as a salt of dicyclohexylamine, cyclohexylamine, sodium, or the like. 4-methoxy-2 obtained in this way,
Histidine having an imidazole group protected by a 3,6-trimethylbenzenesulfonyl group can be very advantageously used in a peptide condensation reaction containing the amino acid residue by conventional means. For example, M. Bodansky and M.A.
Ondetti, Peptide Synthesis
Synthesis), Inter science, New York, 1966
Year: FMFinn and K. Hofmann, The Proteins, Volume 2, H. Neurath, R.
Edited by L. Hiee, Academic Press Inc. New York,
1976; “Peptide Synthesis” by Nobuo Izumiya et al. Maruzen Co., Ltd.
Methods described in 1975, such as the azide method, chloride method, acid anhydride method, mixed acid anhydride method,
DCC method, active ester method, Woodward reagent K
, the carbodiimidazole method, the redox method, and the DCC/HONB method. By the way, in order to perform peptide condensation, α-
When the amino protecting group is a t-butoxycarbonyl group, it is necessary to perform acid treatment with trifluoroacetic acid or the like and then neutralize it with a base such as triethylamine. It is extremely unstable under basic conditions, and even in the case of a para-methoxybenzenesulfonyl group, its stability is insufficient. In comparison, 4-methoxy-2,3,6-trimethylbenzenesulfonyl group is extremely stable, as shown in the examples below, and its usefulness is extremely high. After peptide condensation, the protecting groups of the invention are then removed with acid or 1-hydroxybenzotriazole. As the desorption method using an acid, known acid treatment methods such as anhydrous hydrogen fluoride, methanesulfonic acid, trifluoromethanesulfonic acid, etc. can be applied. Furthermore, in the case of the method of the present invention, as a new acid treatment method,
Trifluoroacetic acid or trifluoroacetic acid containing small amounts of methanesulfonic acid can be used advantageously;
In particular, the elimination action proceeds advantageously when carried out in the presence of thioanisole, anisole or dimethyl sulfide. As described above, the protecting group of the present invention is stable against catalytic reduction and acids such as trifluoroacetic acid, and is also stable against bases such as triethylamine. It has characteristics not found in the base. Next, the present invention will be described with reference examples and examples.
I will explain in more detail. In addition, in this specification, regarding amino acids, peptides, protective groups, active groups, etc., the IUPAC-IUB commission on Biological
It may be indicated by an abbreviation based on nomenclature or by an abbreviation commonly used in the field. Examples of them are shown below. Trp: tryptophan; Lys:
Lysine; His: histidine; Arg: arginine;
Ser: serine; Gly: glycine; Ala: alanine;
Pro: proline; Thr: threonine; Gln: glutamine; Val: valine; Leu: leucine; Ile: isoleucine; Met: methionine; Tyr: tyrosine (unless otherwise specified, amino acids refer to the L-form. However, excluding Gly); Z: carbobenzoxy; Boc: t-butoxycarbonyl;
OBut : t-butyl ester; HONB and
ONB: N-hydroxy-5-norbornene-
2,3-dicarboximide and its esters; HOBt: N-hydroxybenzotriazole; DCC: N,N'-dicyclohexylcarbodiimide; DCU: N,N'-dicyclohexylurea; Mbs: 4-methoxybenzenesulfonyl:
Pme: pentamethylbenzenesulfonyl; Mtr:
4-methoxy-2,3,6-trimethylbenzenesulfonyl; CHA: cyclohexylamine;
DCHA: dicyclohexylamine; DMF: dimethylformamide; TEA: triethylamine;
Further, the developing solvent for thin layer chromatography shown in this specification is Rf 1 : chloroform-methanol-acetic acid (9:1:0.5); Rf 2 : ethyl acetate-pyridine-acetic acid-water (60:20:6: 10); Rf 3 : Chloroform-methanol-water (7:3:0.5); Rf 4 : n
-butanol-pyridine-acetic acid-water (30:20:
6:24). Reference Example 1 (1) Synthesis of 2,3,5-trimethylanisole Dissolve 10 g of 2,3,5-trimethylphenol and 10.4 ml of methyl iodide in 100 ml of dimethyl sulfoxide, cool on ice, and add 60% oil-based 5.6 g of sodium hydride was added and stirred for 10 hours. After adding water to this, it was extracted with ether, and the ether layer was washed with water and dried over anhydrous sodium sulfate. The solvent was evaporated to give an oil. Yield 12.9g (quantitative). (2) Synthesis of 4-methoxy-2,3,6-trimethylbenzenesulfonyl chloride 4.5 g of 2,3,5-trimethylanisole,
After dissolving in 500 ml of methylene chloride and cooling to -5° to -10°C, 400 ml of a methylene chloride solution containing 6.0 ml of chlorosulfonic acid was added dropwise. Thereafter, the temperature was returned to room temperature, and the mixture was poured onto ice containing 5% sodium bicarbonate water. The methylene chloride layer was washed with water and then dried over anhydrous magnesium sulfate. After the solvent was distilled off, crystals were collected by filtration from n-hexane.
Yield 5.0g (67.0%) Melting point 56-58℃ Elemental analysis As C 10 H 13 O 3 SCl Calculated value: C 48.29; H 5.27; S 12.89; Cl 14.26 Experimental value: C 48.42; H 5.21; S 12.61; Cl 14.25 Reference example 2 Production of BOC-His(Mbs)-OH/DCHA 1.50g of BOC-His-OH, 10ml of water, 10ml of acetone
ml of the mixture and cooled on ice. to this
After adding 1.65 ml of TEA, while stirring vigorously, 10 ml of a solution of 1.21 g of Mbs-Cl dissolved in acetone.
was added dropwise, and the mixture was stirred at room temperature for 1 hour. After adding citric acid, the acetone was distilled off, extracted with ethyl acetate, and dried using anhydrous sodium sulfate.
After adding 1.06ml of DCHA and distilling off the solvent,
The residual oil crystallized when left in the refrigerator overnight, so ether was added and this was collected by filtration. Yield 2.65g (74.0%) Melting point 146-147℃ [α] 23 D +21.0゜ (C = 0.93, methanol) Elemental analysis C 30 H 46 O 7 N 4 Calculated value as S: C 59.38; H 7.64; N 9.23; S 5.29 Experimental value: C 59.16; H 7.70; N 8.95; S 5.28 Reference example 3 Production of BOC-His(Mtr)-OH/DCHA 5.11 g of BOC-His-OH was mixed with 30 ml of water and 30 ml of acetone.
ml of the mixture and cooled on ice. to this
After adding 5.60ml of TEA1, while stirring vigorously, 30g of a solution of 4.97g of Mtr-Cl dissolved in acetone.
ml was added dropwise and stirred for 2 hours. After distilling off the acetone, the mixture was acidified with citric acid, extracted with ethyl acetate, and dried over anhydrous sodium sulfate. After adding 3.60ml of DCHA, the solvent was distilled off.
When the residual oil was left in the refrigerator overnight, it crystallized, so ether was added to recrystallize it, and this was collected by filtration. Yield 7.80g (60.1%) Melting point 136−137
°C [α] 23 D +18.8° (C = 1.01, methanol), Rf 1 0.63 Elemental analysis C 21 H 29 O 7 N 3 S・C 12 H 23 N Calculated value: C 61.08; H 8.08; N 8.64 ; S 4.94 Experimental value: C 61.19; H 8.05; N 8.89; S 4.73 Reference example 4 Production of Z-His(Mtr)-OH/DCHA Add 2.90 g of Z-His-OH to 15 ml of water and 15 ml of acetone.
Dissolved in a mixture of and cooled on ice. Add this to 2.80ml of TEA
After adding Mtr−, stirring vigorously,
Drop 15ml of a solution of 2.49g of Cl dissolved in acetone,
I stirred it like that for about an hour. After distilling off the acetone, the residue was acidified with citric acid, extracted with ethyl acetate, and dried using anhydrous sodium sulfate.
After adding 2 ml of DCHA, the solvent was distilled off, ether was added, and the crystals were collected by filtration. Yield 4.10g (60.0%), melting point 149-150℃ [α] 24 D + 14.7゜ (C = 0.87, methanol), Rf 1 0.62 Elemental analysis C 24 H 27 O 7 N 3 S・C 12 H 23 N Calculated value: C 63.32; H 7.38; N 8.21; S 4.07 Experimental value: C 62.94; H 7.11; N 8.11; S 4.80 Test example 4-methoxy-2,3,6-trimethylbenzenesulfonyl group (Mtr) and 4 - Comparison of stability under basic conditions of methoxybenzenesulfonyl group (Mbs) and paratoluenesulfonyl group (Tos) BOC-His (Mtr) -OH・DCHA, BOC-His
(Mbs) −OH・DCHA, BOC−His(Tos)−
Take 100 mg each of OH and DCHA, dissolve in 50% aqueous methanol containing 0.5N-triethylamine, and add 26
After being left at ℃ for a certain period of time, the remaining raw material was quantified using high performance liquid chromatography. As shown in the attached table, the Mtr group can be used with other protecting groups (Mbs,
It is much more stable than Tos), and we were able to confirm its usefulness.
【表】
参考例 ５
Ｈ−His（Mtr）−OHの製造
Ｚ−His（Mtr）−OH・DCHA3.4ｇを酢酸エチ
ル100mlにケンダクしたのち1N−硫酸６mlを加え
てさらに、水を加えてふりまぜたのち、無水硫酸
ナトリウムで乾燥した。溶媒を留去したのち、メ
タノール100mlにとかし、接触還元し、溶媒を留
去すると結晶が析出するので、エーテルを加え
て、これをろ取した。収量1.80ｇ（98.3％）
融点 162−164℃
〔α〕23 D−27.1゜（Ｃ＝1.01、酢酸）、Rf20.35
元素分析 C16H20O5N3S・１／3H2Oとして
計算値：Ｃ 51.60；Ｈ 5.55；Ｎ 11.28
；Ｓ 8.61
実験値：Ｃ 51.76；Ｈ 5.63；Ｎ 11.08
；Ｓ 8.63
実施例 １
鳥GRP（ガストリン・リリージング・ペプチド
の製造
(1) BOC−Leu−MetNH2の製造
BOC−MetNH2（10.3ｇ）に4N−HCl／酢酸40
mlを加えたのち、振りまぜると結晶が析出するの
でエーテルを加えて、これをろ取し、乾燥した。
これをDMF200mlにとかし、氷冷し、TEA7.0ml
を加えて中和した。これにBOC−Leu−ONB
（BOC−Leu−OH8.0ｇ，HONB6.85ｇ，
DCC7.83ｇより調製する）を加えて、一夜かきま
ぜた。溶媒を留去したのち酢酸エチルにとかし、
重曹水、クエン酸水で洗浄したのち、無水硫酸ナ
トリウムを用いて乾燥した。溶媒を留去すると結
晶が析出するので、エーテルを加えて、これをろ
取し、メタノール−エーテルより再結晶した。収
量10.8ｇ（86.3％）
融点 152−154℃、〔α〕23 D−34.2゜（Ｃ＝1.03，
DMF）、Rf10.65
元素分析 C16H31O4N3Sとして
計算値：Ｃ 53.15；Ｈ 8.64；Ｎ 11.63
；Ｓ 8.87
実験値：Ｃ 53.56；Ｈ 8.72；Ｎ 11.47
；Ｓ 8.92
(2) BOC His（Mtr）OH・DCHAの製造
BOC−His OH5.11ｇを水30ml、セトン30mlの
混合液にとかし、氷冷した。これに、TEA5.6ml
を加えたのち、Mtr−Cl4.97ｇのアセトン溶液30
mlを加え、２時間かきまぜた。アセトンを減圧留
去したのち、クエン酸酸性として酢酸エチルで抽
出し、水洗したのち、無水硫酸ナトリウムを用い
て乾燥した。溶媒を留去したのち、少量の酢酸エ
チルにとかし、DCHA3.6mlを加え、溶媒を留去
して冷蔵庫内で一夜放置した。析出した結晶にエ
ーテルを加えて、これをろ取した。収量7.80ｇ
（60.1％）
融点 136−137℃、〔α〕23 D＋18.8゜（ｃ＝1.01、
メタノール）、Rf10.63
元素分析 C33H52O7N4Sとして
計算値：Ｃ 61.08；Ｈ 8.08；Ｎ 8.64
；Ｓ 4.94
実験値：Ｃ 61.19；Ｈ 8.05；Ｎ 8.89
；Ｓ 4.73
(3) BOC−His（Mtr）−Leu−MetNH2の製造
BOC−Leu−MetNH210.0ｇに、4N−HCl／酢
酸30mlを加え、室温で20分間放置したのちエーテ
ルを加えて沈澱としてろ取し乾燥した。一方
BOC−His（Mtr）OH・DCHA16.0ｇを、酢酸エ
チルに懸濁し、1N−硫酸27mlを加えて分液し、
無水硫酸ナトリウムを用いて乾燥した。溶媒を留
去したのち、アセトニトリル100mlにとかし、
HONB5.0ｇを加えて氷冷し、DCC5.8ｇを加え
て、一夜反応したのち、析出するDCUをろ去し
た。先に調製したアミン成分をDMF100mlにとか
し、TEA4.6mlで中和したのち、活性エステルを
加えて、６時間かきまぜた。溶媒を留去したの
ち、酢酸エチルに転溶し、重曹水、クエン酸水、
水の順に洗浄し、無水硫酸ナトリウムで乾燥し
た。溶媒を留去したのち、エーテルを加えて粉末
としてろ取した。
収量 15.1ｇ（86.0％）
融点 129−131℃、〔α〕23 D−13.8゜（ｃ＝0.87，
DMF）、Rf10.64
元素分析 C32H50O8N6S2として
計算値：Ｃ 54.06；Ｈ 7.09；Ｎ 11.82
；Ｓ 9.02
実験値：Ｃ 54.69；Ｈ 7.39；Ｎ 11.97
；Ｓ 8.39
(4) Ｚ−Val−Gly−OButの製造
Ｚ−Gly−OBut12.0ｇをメタノール中接触還元
したのち溶媒を留去し、DMFにとかし氷冷した。
これに、Ｚ−Val−OH8.80ｇ，HONB7.20ｇ，
DCC8.24ｇを加え、一夜かきまぜた。析出した
DCUをろ去したのち残留物を酢酸エチルにとか
し、重曹水、クエン酸水で洗浄したのち、無水硫
酸ナトリウムで乾燥した。溶媒を留去し、残留物
に石油ベンジンを加えて結晶としてろ取したの
ち、酢酸エチル−石油ベンジンより再結晶した。
収量11.1ｇ（87.0％）
融点 141℃，〔α〕23 D−2.1゜（ｃ＝1.16，DMF）、
Rf10.80
元素分析 C19H28O5N2として
計算値：Ｃ 62.62；Ｈ 7.74；Ｎ 7.69
実験値：Ｃ 62.49；Ｈ 7.60；Ｎ 7.72
(5) Ｚ−Ala−Val−Gly−OButの製造
Ｚ−Val−Gly−OBut10.0ｇをメタノール中、
接触還元したのち、DMF100mlにとかし、Ｚ−
Ala−OH5.7ｇ，HONB4.1ｇを加え、氷冷した。
これにDCC6.2ｇを加えて、一夜かきまぜた。析
出したDCUをろ去し、溶媒を留去すると、結晶
が析出するので、酢酸エチルを加えて、結晶をろ
取し、よく洗浄した。
収量 10.5ｇ（96.4％）
融点 184−185℃，〔α〕23 D−7.8゜（ｃ＝1.06，
DMF）、Rf10.67
元素分析 C22H33O6N3として
計算値：Ｃ 60.67；Ｈ 7.64；Ｎ 9.65
実験値：Ｃ 60.93；Ｈ 7.86；Ｎ 9.77
(6) Ｚ−Trp（Mtr）−Ala−Val−Gly−OButの
製造
Ｚ−Ala−Val−Gly−OBut5.0ｇをメタノール
中、接触還元したのち、溶媒を留去し、DMF100
mlにとかした。これに、Ｚ−Trp（Mtr）OH6.34
ｇ，HOBT1.86ｇを加えて氷冷し、DCC2.85ｇを
加え、一夜かきまぜた。析出したDCUをろ去し、
溶媒を留去したのち、少量のｎ−ブタノールを含
む酢酸エチルで抽出し重曹水で洗浄したのち、無
水硫酸ナトリウムで乾燥した。溶媒を留去し、エ
ーテルを加えて沈澱としてろ取したのち、メタノ
ール−酢酸エチル−エーテルより結晶として、こ
れをろ取した。
収量 8.80ｇ（96.6％）
融点 154−155℃，〔α〕23 D−22.7゜（ｃ＝0.88，
DMF）、Rf10.68
元素分析 C43H56O10N5Sとして
計算値：Ｃ 61.85；Ｈ 6.76；Ｎ 8.39
；Ｓ 3.84
実験値：Ｃ 62.06；Ｈ 7.01；Ｎ 8.58
；Ｓ 3.58
(7) BOC−His（Mtr）−Trp（Mtr）−Ala−Val−
Gly−OButの製造
Ｚ−Trp（Mtr）−Ala−Val−GlyOBut8.0ｇを
DMF−メタノール中、接触還元したのち、メタ
ノールを留去した。これにBOC−His（Mtr）OH
（BOC−His（Mtr）OH・DCHA6.22ｇより調
製）、HONB1.90ｇを加えて氷冷したのち、
DCC2.18ｇを加え、一夜かきまぜた。析出した
DCUをろ去したのち、溶媒を留去し、エーテル
を加えて、沈澱としてろ取した。これをメタノー
ル、酢酸エチル、エーテルの混合液で洗浄した。
収量10.2ｇ（92.5％）
融点 202−204℃，〔α〕23 D−16.1゜（ｃ＝1.18，
DMF）、Rf10.68
元素分析 C56H77O14N8S2として
計算値：Ｃ 58.46；Ｈ 6.75；Ｎ 9.74
；Ｓ 5.57
実験値：Ｃ 58.32；Ｈ 6.54；Ｎ 9.58
；Ｓ 5.51
(8) BOC−Ser−His（Mtr）−Trp（Mtr）−Ala−
Val−GlyOHの製造
BOC−His（Mtr）−Trp（Mtr）−Ala−Val−
GlyOBut4.0ｇにTFA30mlを加え、室温で50分間
振りまぜたのち、TFAを留去し、エーテルを加
えて、沈澱としろ取し、乾燥した。一方BOC−
Ser−OH0.76ｇ，HONB0.72ｇをアセトニトリル
20mlにとかし、氷冷しこれにDCC0.83ｇを加えて
４時間かきまぜた。先に調製したアミン成分を
DMF50mlにとかし、TEA1.0mlを加えて中和し
たのち、活性エステルを加えて、一夜かきまぜ
た。溶媒を留去したのち、少量の酢酸を加えて、
さらに水を加えて沈澱としてろ取した。これを
DMF−水より再沈でんした。収量 3.60ｇ（85.0
％）
融点 148−152℃，〔α〕23 D−16.4゜（ｃ＝1.09，
DMF）
元素分析 C55H74O16N9S2・2H2Oとして
計算値：Ｃ 54.26；Ｈ 6.46；Ｎ 10.36
；Ｓ 5.27
実験値：Ｃ 54.58；Ｈ 6.33；Ｎ 10.82
；Ｓ 5.51
(9) Ｚ−Arg（Pme）−Gly−OButの製造
Ｚ−Gly・OBut13ｇをメタノール300ml中、接
触還元したのち、溶媒を留去し、残留物を
DMF200mlにとかした。これに、Ｚ−Arg（Pme）
OH〔Ｚ−Arg（Pme）OH・CHA20ｇより調製〕
を加え、氷冷し、HOBT5.4ｇ，DCC8.2ｇを加え
て一夜かきまぜた。析出したDCUをろ去したの
ち、溶媒を留去し、残留物を酢酸エチルにとかし
た。これを、重曹水、クエン酸水で洗浄したの
ち、無水硫酸ナトリウムで乾燥した。溶媒を留去
したのち、石油ベンジンを加えて粉末としてろ取
した。収量19.8ｇ（95.0％）
融点 55−60℃，〔α〕23 D＋0.2゜（ｃ＝0.88，
DMF）、Rf10.62
元素分析 C31H45O7N5Sとして
計算値：Ｃ 58.93；Ｈ 7.18；Ｎ 11.09
；Ｓ 5.08
実験値：Ｃ 58.96；Ｈ 7.01；Ｎ 10.67
；Ｓ 5.05
(10) Ｚ−Tyr−Pro−OButの製造
Ｚ−Pro OBut15.0ｇをメタノール300mlにとか
し接触還元したのち、溶媒を留去し、DMF400ml
にとかした。これにＺ−Tyr−OH（Ｚ−Tyr−
OH・DCHA 20.0ｇより調製）、HOBT6.75ｇを
加えて氷冷し、さらにDCC 10.4ｇを加えて、一
夜かきまぜた。析出したDCUをろ去し、溶媒を
留去したのち、酢酸エチルにとかし、重曹水、ク
エン酸水で洗浄し、無水硫酸ナトリウムで乾燥し
た。溶媒を留去したのち、シリカゲルのカラム
（7.5×９cm）に付し、１％（MeOH／クロロホル
ムで溶出し、目的物の分画を集め、濃縮したの
ち、石油ベンジンを加えて粉末としてろ取した。
収量15.4ｇ（82.2％）
〔α〕23 D−39.9゜（ｃ＝0.83，DMF）Rf10.62
元素分析 C26H32O6N2・1/2H2Oとして
計算値：Ｃ 65.39；Ｈ 6.97；Ｎ 5.87
実験値：Ｃ 65.70；Ｈ 6.93；Ｎ 5.66
(11) Ｚ−Ile−Tyr−Pro−OBut
Ｚ−Tyr−Pro−OBut15.2ｇをメタノール300
mlにとかし、接触還元したのち、溶媒を留去し、
残留物をDMF200mlにとかした。これにＺ−
IleOH8.0ｇ，HONB6.5ｇを加え氷冷したのち、
DCC7.4ｇを加えて、一夜かきまぜた。少量のＮ，
Ｎ−ジメチルプロパンジアミンを加えたのち、
DCUをろ去し、溶媒を減圧で留去した。残留物
を酢酸エチルにとかしたのち、重曹水、クエン酸
水で洗浄し、無水硫酸ナトリウムで乾燥した。溶
媒を留去すると結晶が析出するので、石油ベンジ
ンを加えて、これをろ取したのち、メタノール、
エーテル、石油ベンジンより再結晶した。収量
10.9ｇ（62.5％）
融点 177−178℃，〔α〕23 D−38.3゜（ｃ＝1.11，
DMF）、Rf10.62
元素分析 C32H43O7N3として
計算値：Ｃ 66.07；Ｈ 7.45；Ｎ 7.22
実験値：Ｃ 66.07；Ｈ 7.74；Ｎ 7.19
(12) Ｚ−Ile−Try−Pro−OHの製造
Ｚ−Ile−Tyr−Pro OBut6.0ｇにTFA60mlを
加え、室温で１時間ふりまぜたのち、留去し、残
留物を酢酸エチルにとかし、水洗したのち無水硫
酸ナトリウムで乾燥した。溶媒を留去したのちエ
ーテルを加えて粉末として、ろ取した。
収量 5.10ｇ（94.2％）
融点 72−74℃，〔α〕23 D−25.2゜（ｃ＝1.01，
DMF）、Rf10.44
元素分析 C28H35O7N3として
計算値：Ｃ 63.98；Ｈ 6.71；Ｎ 7.99
実験値：Ｃ 63.75；Ｈ 6.67；Ｎ 7.84
(13) Ｚ−Ile−Tyr−Pro−Arg（Pme）−
GlyOButの製造
Ｚ−Arg（Pme）−Gly−OBut7.58ｇをメタノー
ル300mlにとかし、1N−塩酸12mlを加えたのち、
接触還元した。溶媒を留去したのち残留物を
DMF100mlにとかし、氷冷して、TEA2.00mlを
加えて中和した。これに、Ｚ−Ile−Tyr−Pro−
OH5.78ｇ，HOBT2.23ｇ，DCC3.40ｇを加えて、
一夜かきまぜた。析出したDCUをろ去したのち、
溶媒を留去し、残留物を酢酸エチルにとかした。
これを、重曹水、クエン酸水で洗浄し、無水硫酸
ナトリウムで乾燥した。溶媒を留去し、残留物を
シリカゲルのカラム（5.5×10cm）に付し、２％
MeOH／クロロホルムで溶出し、目的物の分画
を集め濃縮したのち、エーテルを加えて、粉末と
してろ取した。
収量 7.20ｇ（62.2％）
融点 110−112℃，〔α〕23 D−28.3゜（ｃ＝1.19，
DMF）、Rf10.59
元素分析 C51H72O11N8S・H2Oとして
計算値：Ｃ 59.85；Ｈ 7.29；Ｎ 10.93；Ｓ
3.13
実験値：Ｃ 60.16；Ｈ 7.56；Ｎ 10.95；Ｓ
2.98
(14) Ｚ−Lys（Mtr）−Ile−Tyr−Pro−Arg
（Pme）−GlyOButの製造
Ｚ−Ile−Tyr−Pro−Arg（Pme）−Gly−OBut
7.0ｇをメタノール350mlにとかし、1N−塩酸７
mlを加えて、接触還元した。溶媒を留去したの
ち、残留物をDMFを200mlにとかし、氷冷し、
TEA1.0mlを加えて中和した。これに、Ｚ−Lys
（Mtr）−OH〔Ｚ−Lys（Mtr）OH・DCHA4.74ｇ
より調製〕、HOBT120ｇ，DCC1.80ｇを加えて、
一夜かきまぜた。析出したDCUをろ去し残留物
を酢酸エチルにとかした。これを重曹水、クエン
酸水で洗浄したのち、無水硫酸ナトリウムで乾燥
した。溶媒を留去し、エーテルを加えて粉末とし
たのち、酢酸エーテル−エーテルより２回再沈澱
した。収量8.40ｇ（87.5％）
融点 116〜118℃，〔α〕23 D−19.9゜（ｃ＝1.05，
DMF）、Rf10.59
元素分析 C66H96O15N10S2として
計算値：Ｃ 59.44；Ｈ 7.26；Ｎ 10.50
；Ｓ 4.81
実験値：Ｃ 59.10；Ｈ 7.44；Ｎ 10.46
；Ｓ 4.98
(15) Ｚ−Thr−Lys（Mtr）−Ile−Tyr−Pro−
Arg（Pme）−Gly−OButの製造
Ｚ−Lys（Mtr）−Ile−Tyr−Pro−Arg（Pme）
−GlyOBut7.50ｇをメタノール350ml中、接触還
元したのち、溶媒を留去し、これをDMF100mlに
とかした。これに、Ｚ−Thr−OH1.49ｇ，
HONB1.51ｇを加えて、氷冷したのち、DCC1.73
ｇを加えて一夜かきまぜた。少量のＮ，Ｎ−ジメ
チルプロパンジアミンを加え、DCUをろ去した
のち、溶媒を留去し、残留物を少量のｎ−ブタノ
ールを含む酢酸エチルにとかした。これを重曹水
で洗浄したのち、水洗し無水硫酸ナトリウムで乾
燥した。溶媒を留去し、残留物にエーテルを加え
て、粉末としたのち、これを、ろ取した。収量
8.0ｇ（99.2％）
融点 122−124℃，〔α〕23 D−25.5゜（ｃ＝0.82，
DMF）、Rf10.59
元素分析 C70H103O17N11S2として
計算値：Ｃ 58.60；Ｈ 7.24；Ｎ 10.74；Ｓ
4.47
実験値：Ｃ 58.61；Ｈ 7.29；Ｎ 10.47；Ｓ
4.02
(16) BOC−Leu−Thr−Lys（Mtr）−Ile−Tyr−
Pro−Arg（Pme）−GlyOButの製造
Ｚ−Thr−Lys（Mtr）−Ile−Tyr−Pro−Arg
（Pme）−GlyOBut7.5ｇをメタノール350mlにとか
し、パラトルエンスルホン酸0.99ｇを加えて、接
触還元したのち、溶媒を留去した。これを
DMF100mlにとかし、氷冷し、TEA0.74mlを加
えて中和した。これに、BOC−Leu−OH1.37ｇ，
HONB1.41ｇ，DCC1.62ｇを加えて一夜かきまぜ
た。DCUをろ去し、溶媒を留去したのち、酢酸
エチルにとかし、重曹水、クエン酸水で洗浄し無
水硫酸ナトリウムで乾燥した。溶媒を留去したの
ち、エーテルを加えて、粉末としてろ取した。収
量7.55ｇ（95.4％）
融点 136−138℃，〔α〕23 D−26.6゜（ｃ＝1.00，
DMF）、Rf10.59
元素分析 C73H116O18N12S2・H2Oとして
計算値：Ｃ 57.23；Ｈ 7.76；Ｎ 10.97
；Ｓ 4.19
実験値：Ｃ 57.53；Ｈ 8.12；Ｎ 10.65
；Ｓ 3.90
(17) BOC−Ala−Leu−Thr−Lys（Mtr）−Ile−
Tyr−Pro−Arg（Pme）−Gly−OHの製造
BOC−Leu−Thr−Lys（Mtr）−Ile−Tyr−Pro
−Arg（Pme）−Gly−OBut4.20ｇにTFA40mlを加
え、室温で60分間ふりまぜたのち溶媒を留去し、
エーテルを加えて粉末としてろ取し、乾燥した。
これをDMF50mlにとかし、氷冷し、TEA0.80ml
を加えた。これに、BOC−Ala−ONB（BOC−
Ala−OH0.52ｇ，HONB0.54ｇ，DCC0.62ｇよ
り調製）を加えて、一夜かきまぜた。少量のＮ，
Ｎ−ジメチルプロパンジアミンを加えて溶媒を留
去したのち、酢酸水を加えて沈澱としてろ取し
た。これを、含水メタノールで再沈澱した。収量
3.60ｇ（88.1％）
融点 128−130℃，〔α〕23 D−32.9゜（ｃ＝0.98，
DMF），Rf10.40
元素分析 C72H113O19N13S2・2H2Oとして
計算値：Ｃ 55.26；Ｈ 7.54；Ｎ 11.64
；Ｓ 4.10
実験値：Ｃ 55.02；Ｈ 6.89；Ｎ 11.38
；Ｓ 3.77
(18) Ｚ−Ser−ProOButの製造
Ｚ−Pro−OBut11.0ｇをメタノール300mlにと
かし、接触還元したのち、溶媒を留去し、
DMF200mlにとかした。これにＺ−Ser−OH7.2
ｇ，HOBT4.90ｇ，DCC7.50ｇを加えて、一夜か
きまぜた。析出したDCUをろ去し、溶媒を留去
し、残留物を酢酸エチルにとかした。これを、重
曹水、クエン酸水で洗浄したのち、無水硫酸ナト
リウムで乾燥した。溶媒を留去し、石油ベンジン
を加えて、結晶としてろ取したのち、酢酸エチル
−エーテルより再結晶した。
収量 9.50ｇ（67.2％）
融点 126−127℃，〔α〕23 D−50.9゜（ｃ＝0.95，
DMF）、Rf10.65
元素分析 C20H28O6N2として
計算値：Ｃ 61.21；Ｈ 7.19；Ｎ 7.14
実験値：Ｃ 61.45；Ｈ 7.16；Ｎ 7.31
(19) Ｚ−Gly−Ser−Pro−OButの製造
Ｚ−Ser−Pro−OBut10.0ｇをメタノール300ml
にとかし、接触還元したのち、溶媒を留去し、
DMF300mlにとかした。これにＺ−Gly−OH5.06
ｇ，HONB5.13ｇを加えて氷冷したのち、
DCC5.89ｇを加えて、一夜かきまぜた。少量の
Ｎ，Ｎ−ジメチルプロパンジアミンを加えたの
ち、DCUをろ去し、溶媒を留去した。残留物を
酢酸エチルにとかし、重曹水、クエン酸水で洗浄
し、無水硫酸ナトリウムで乾燥した。溶媒を留去
し、石油ベンジンを加えて結晶としてろ取したの
ち、エーテルで再結晶した。収量7.70ｇ（72.1
％）
融点 96−98℃，〔α〕23 D−53.4゜（ｃ＝1.05，
DMF）、Rf10.61
元素分析 C22H31O7N3として
計算値：Ｃ 58.78；Ｈ 6.95；Ｎ 9.35
実験値：Ｃ 58.86；Ｈ 7.04；Ｎ 9.46
(20) Ｚ−Gly−Gly−Ser−Pro−OButの製造
Ｚ−Gly−Ser−Pro−OBut7.0ｇをメタノール
200mlにとかし、接触還元したのち、溶媒を留去
し、残留物をDMF100mlにとかした。これにＺ−
Gly−OH3.0ｇ，HONB3.10ｇを加えて氷冷した
のち、DCC3.60ｇを加えて一夜かきまぜた。少量
のＮ，Ｎ−ジメチルプロパンジアミンを加えたの
ち、DCUをろ去し、溶媒を留去した。残留物を
酢酸エチルにとかし、飽和食塩水で洗浄したの
ち、無水硫酸ナトリウムで乾燥した。溶媒を留去
しエーテルを加えて結晶としたのちろ取し、酢酸
エチルで再結晶した。収量5.70ｇ（78.5％）
融点 129−130℃，〔α〕23 D−47.0゜（ｃ＝0.81，
DMF）、Rf10.47
元素分析 C24H34O8N4として
計算値：Ｃ 56.90；Ｈ 6.77；Ｎ 11.06
実験値：Ｃ 56.75；Ｈ 6.68；Ｎ 10.90
(21) Ｚ−Gln−Pro−OButの製造
Ｚ−Pro−OBut16.2ｇをメタノール350mlにと
かし、接触還元したのち溶媒を留去し、DMF200
mlにとかした。これにＺ−Gln−OH12.4ｇ，
HOBT7.16ｇを加えて、氷冷し、DCC10.9ｇを加
えて一夜かきまぜた。析出したDCUをろ去し、
溶媒を留去したのち、酢酸エチルにとかし、重曹
水、クエン酸水で洗浄し、無水硫酸ナトリウムで
乾燥した。溶媒を留去したのち、石油ベンジンを
加えて結晶としてこれをろ取し、酢酸エチル−石
油ベンジンより再結晶した。
収量 15.8ｇ（82.5％）
融点 106−107℃，〔α〕23 D−51.2゜（ｃ＝1.06，
DMF）、Rf10.62
元素分析 C22H31O6N3として
計算値：Ｃ 60.95；Ｈ 7.21；Ｎ 9.69
実験値：Ｃ 60.95；Ｈ 7.36；Ｎ 9.41
(22) Ｚ−Leu−Gln−Pro−OButの製造
Ｚ−Gln−Pro−OBut8.0ｇ、パラトルエンスル
ホン酸3.51ｇをメタノール300mlにとかし接触還
元したのち、溶媒を留去した。残留物をDMF200
mlにとかしたのち氷冷し、TEA2.6mlを加えて中
和した。これにＺ−LeuOH（Ｚ−Leu−OH・
DCHA8.24ｇより調製）、HONB4.00ｇ，
DCC4.60ｇを加えて一夜かきまぜた。少量のＮ，
Ｎ−ジメチルプロパンジアミンを加えたのち
DCUをろ去し、溶媒を留去した。残留物を酢酸
エチルにとかし、重曹水、クエン酸水で洗浄し、
無水硫酸ナトリウムで乾燥した。溶媒を留去し結
晶を析出させたのち、石油ベンジンを加えて、こ
れをろ取し、酢酸エチル−石油ベンジンで再結晶
した。収量7.70ｇ（76.3％）
融点 62−64℃，〔α〕23 D−51.7゜（ｃ＝1.11，
DMF）、Rf10.61
元素分析 C28H42O7N4として
計算値：Ｃ 61.52；Ｈ 7.75；Ｎ 10.25
実験値：Ｃ 61.19；Ｈ 7.75；Ｎ 10.11
(23) BOC−Pro−Leu−Gln−Pro−OButの製造
Ｚ−Leu−Gln−Pro−OBut7.0ｇを、メタノー
ル300mlにとかし、接触還元したのち、溶媒を留
去し、残留物をDMF100mlにとかした。これに
BOC−ProOH2.42ｇ，HONB2.76ｇを加えて氷
冷したのち、DCC3.17ｇを加えて、一夜かきまぜ
た。少量のＮ，Ｎ−ジメチルプロパンジアミンを
加えたのち、DCUをろ去し、溶媒を留去し、酢
酸エチルにとかした。これを、重曹水、クエン酸
水で洗浄したのち、無水硫酸ナトリウムで乾燥し
た。溶媒を留去したのち、エーテル−石油ベンジ
ンより粉末として、ろ取した。収量6.50ｇ（83.3
％）
融点 74−76℃，〔α〕23 D−76.3゜（ｃ＝1.19，
DMF）、Rf10.60
元素分析 C30H51O8N5として
計算値：Ｃ 59.05；Ｈ 8.43；Ｎ 11.49
実験値：Ｃ 58.89；Ｈ 8.12；Ｎ 11.08
(24) BOC−Ala−Pro−Leu−Gln−Pro−OHの
製造
BOC−Pro−Leu−Gln−Pro−OBut6.0ｇに
TFA60mlを加え、室温で60分間ふりまぜたのち、
溶媒を留去し、残留物にエーテルを加えて粉末と
して、ろ取し乾燥した。これをDMF100mlにとか
し氷冷し、TEA2.80mlを加えたのち、BOC−Ala
−ONB（BOC−Ala−OH1.95ｇ，HONB2.07ｇ，
DCC2.38ｇより調製）を加えて一夜かきまぜた。
溶媒を留去したのち、少量の酢酸を加え、次いで
エーテルを加えて粉末としてろ取した。これをク
ロロホルムにとかし、シリカゲルのカラム（5.5
×８cm）に付し、５％メタノール／クロロホルム
で溶出し、目的物の分画を取り、濃縮したのち、
エーテルを加えて粉末としてろ取した。収量4.50
ｇ（73.2％）
融点 124−128℃，〔α〕23 D−82.1゜（ｃ＝1.04，
DMF）、Rf10.18
元素分析 C29H48O9N6として
計算値：Ｃ 55.75；Ｈ 7.74；Ｎ 13.45
実験値：Ｃ 55.38；Ｈ 7.65；Ｎ 13.21
(25) BOC−Ala−Pro−Leu−Gln−Pro−Gly−
Gly−Ser−Pro−OButの製造
Ｚ−Gly−Gly−Ser−Pro−OBut2.23ｇをメタ
ノール100mlにとかし接触還元したのち、溶媒を
留去し、残留物をDMF50mlにとかした。これに
BOC−Ala−Pro−Leu−Gln−Pro−OH2.50ｇ，
HONB0.90ｇを加えて氷冷し、さらにDCC1.03ｇ
を加えて一夜かきまぜた。析出したDCUをろ去
したのち溶媒を留去し、これを少量のｎ−BuOH
を含む酢酸エチルにとかし、飽和食塩水で洗浄し
たのち無水硫酸ナトリウムで乾燥した。溶媒を留
去したのち、エーテルを加えて粉末としてろ取し
た。収量3.85ｇ（98.3％）
融点 100−105℃，〔α〕23 D−74.0゜（ｃ＝0.92，
DMF）、Rf10.19
元素分析 C45H74O14N10・H2Oとして
計算値：Ｃ 54.20；Ｈ 7.68；Ｎ 14.05
実験値：Ｃ 54.01；Ｈ 7.45；Ｎ 13.44
(26) BOC−Ala−Pro−Leu−Gln−Pro−Gly−
Gly−Ser−Pro−OHの製造
BOC−Ala−Pro−Leu−Gln−Pro−Gly−Gly
−Ser−Pro−OBut1.0ｇにTFA10mlを加え、室
温で１時間ふりまぜたのち溶媒を留去し、エーテ
ルを加えて粉末としてろ取した。これをDMF10
mlにとかし、氷冷したのちTEA0.46mlを加え、
次にBOC−OH0.27ｇを加えて４時間かきまぜ
た。溶媒を留去したのち、酢酸1.5mlを加え、エ
ーテルを加えて粉末としてろ取した。これをメタ
ノール−エーテルより再沈澱した。収量0.87ｇ
（92.4％）
融点141−145℃，〔α〕23 D−72.9゜（ｃ＝0.98，
DMF）、Rf30.29
元素分析 C41H66O14N10・3H2Oとして
計算値：Ｃ 50.40；Ｈ 7.43；Ｎ 14.34
実験値：Ｃ 50.16；Ｈ 6.76；Ｎ 14.11
(27) BOC−Ser−His（Mtr）−Trp（Mtr）−Ala
−Val−Gly−His（Mtr）−Leu−Met−NH2の
製造
BOC−His（Mtr）−Leu−MetNH23.97ｇに
TFA40mlを加え、室温で10分間ふりまぜたのち、
1.3N−塩酸4.1mlを加えて溶媒を留去し、エーテ
ルを加えて粉末としてろ取し、乾燥した。これを
DMF40mlにとかし、氷冷し、TEA0.86mlを加
え、次いでBOC−Ser−His（Mtr）−Trp（Mtr）−
Ala−Val−Gly−OH5.50ｇ，HONB1.0ｇ，
DCC1.15ｇを加え一夜かきまぜた。析出した
DCUをろ去し、溶媒を留去したのち、水を加え
て、粉末としてろ取した。次いでこれを含水エタ
ールで洗浄した。
収量 7.35ｇ（88.9％）
融点 192−193℃，〔α〕23 D−12.5゜（ｃ＝1.05，
DMF）、Rf10.50
元素分析 C82H114O21N15S4として
計算値：Ｃ 55.51；Ｈ 6.48；Ｎ 11.84
；Ｓ 7.23
実験値：Ｃ 55.56；Ｈ 6.74；Ｎ 11.83
；Ｓ 6.55
(28) BOC−Ser−His−Trp（Mtr）−Ala−Val
−Gly−His−Leu−Met−NH2の製造
BOC−Ser−His（Mtr）−Trp（Mtr）−Ala−
Val−Gly−His（Mtr）−Leu−Met−NH23.0ｇを
DMF15mlにとかし、HOBT2.30ｇを加えて30分
間処理したのち、溶媒を留去し、エーテルを加え
て粉末としてろ取した。収量2.15ｇ（92.5％）
融点 191−193℃，〔α〕23 D−18.8゜（ｃ＝0.94，
DMF）、Rf30.64
元素分析 C62H90O15N15S2・H2Oとして
計算値：Ｃ 54.45；Ｈ 6.78；Ｎ 15.36
；Ｓ 4.69
実験値：Ｃ 53.92；Ｈ 6.43；Ｎ 15.80
；Ｓ 4.34
(29) BOC−Ala−Leu−Thr−Lys（Mtr）−Ile−
Tyr−Pro−Arg（Pme）−Gly−Ser−His−
Trp（Mtr）−Ala−Val−Gly−His−Leu−Met
−NH2の製造
BOC−Ser−His−Trp（Mtr）−Ala−Val−
Gly−His−Leu−MetNH22.0ｇにTFA20mlを加
え、室温で15分間処理したのち留去し、エーテル
を加えて粉末としてろ取し、乾燥した。これを
DMF5mlにとかし、TEA1.24mlを加えてよくか
きまぜたのち、エーテルを加えて沈澱させ、粉末
としてろ取した。これをエーテル20mlにとかし、
BOC−Ala−Leu−Thr−Lys（Mtr）−Ile−Tyr−
Pro−Arg（Pme）−Gly−OH1.78ｇ，HONB0.41
ｇを加えたのち氷冷し、DCC0.47ｇを加えて一夜
かきまぜた。溶媒を留去したのち、エタノール−
酢酸エチルを加えて粉末としてろ取したのち、熱
エタノールで洗浄した。
収量 3.30ｇ（99.7％）
融点 222−223℃（分解），〔α〕23 D−22.8゜（ｃ
＝
1.02，DMF）、Rf30.67
元素分析 C129H193O31N28S4・8H2Oとして
計算値：Ｃ 53.34；Ｈ 7.25；Ｎ 13.50
；Ｓ 4.42
実験値：Ｃ 53.35；Ｈ 6.98；Ｎ 12.91
；Ｓ 4.58
(30) BOC−Ala−Pro−Leu−Gln−Pro−Gly−
Gly−Ser−Pro−Ala−Leu−Thr−Lys（Mtr）
−Ile−Tyr−Pro−Arg（Pme）Gly−Ser−His
（Mtr）−Trp（Mtr）−Ala−Val−Gly−His−
Leu−Met−NH2の製造
BOC−Ala−Leu−Thr−Lys（Mtr）−Ile−Tyr
−Pro−Arg（Pme）−Gly−Ser−His−Trp
（Mtr）−Ala−Val−Gly−His−Leu−Met−
NH2500mgに、TFA5mlを加え、ふりまぜたのち、
留去し、エーテルを加えて粉末としてろ取した。
これをDMF1mlにとかし、TEA0.2mlを加えて、
よくかきまぜたのち、エーテルを加えて粉末とし
てろ取した。これを、DMF5mlにとかし、BOC
−Ala−Pro−Leu−Gln−Pro−Gly−Gly−Ser
−Pro−OH217mg，HOBT50mgを加えたのち氷
冷し、DCC160mgを加えて一夜かきまぜた。析出
したDCUをろ去したのち、溶媒を留去し、残留
物に酢酸エチルを加えて粉末としてろ取した。こ
れをDMF−メタノールにとかし約30分間加熱し
たのち、メタノールを留去し、残留物に酢酸エチ
ルを加えて粉末としてろ取した。次いでこれをメ
タノール水で洗浄した。収量480mg（72.2％）
融点 203−208℃（分解），〔α〕23 D−35.2゜（ｃ
＝
0.93，DMF）、Rf30.66
元素分析 C165H249O42N38S4・6H2Oとして
計算値：Ｃ 53.95；Ｈ 7.16；Ｎ 14.49
；Ｓ 3.49
実験値：Ｃ 54.08；Ｈ 6.87；Ｎ 14.19
；Ｓ ３，43
(31) Ｈ−Ala−Pro−Leu−Gln−Pro−Gly−
Gly−Ser−Pro−Ala−Leu−Thr−Lys−Ile
−Tyr−Pro−Arg−Gly−Ser−His−Trp−
Ala−Val−Gly−His−Leu−Met−NH2（鳥
GRP）の製造
BOC−Ala−Pro−Leu−Gln−Pro−Gly−Gly
−Ser−Pro−Ala−Leu−Thr−Lys（Mtr）−Ile
−Tyr−Pro−Arg（Pme）−Gly−Ser−His−
Trp（Mtr）−Ala−Val−Gly−His−Leu−Met−
NH2100mgに0.3Mメタンスルホン酸／TFA−チ
オアニソール（９：１）８mlを加え、室温で２時
間ふりまぜたのち、酢酸アンモニア100mgを加え
て、TFAを留去し、エーテルを加えて粉末とし
てろ取した。これを、少量の1N酢酸水にとかし、
セフアデツクスＧ−25のカラム（2.2×120cm）に
付した。1N−酢酸で溶出し170ml−270mlの分画
を取り、凍結乾燥したのち、少量の水にとかし
Amberlite IRA410（１×10cm）のカラムを通し
たのち、カルボキシメチルセルロースのカラム
（2.2×17cm）に付した。水（400ml）−0.4M酢酸
アンモニア水（400ml）の線型勾配法で溶出し、
335〜365mlの分画を集めて凍結乾燥した。収量28
mg（35％）。
〔α〕22 D−102.2゜（ｃ＝0.32，１％酢酸）Rf40.39
アミノ酸分析値（４％チオグリコール酸／6N
塩酸加水分解）：Lys1.00(1)；His1.73(2)；Arg1.04
(1)；Trp0.67(1)；Thr1.06(1)；Ser1.77(2)；Glu1.11
(1)；Pro4.20(4)；Gly4.23(4)；Ala3.19(3)；Val1.06
(1)；Met1.02(1)；Ile0.96(1)；Leu3.04(3)；Tyr1.00
(1)（平均回収率73.7％）[Table] Reference Example 5 Production of H-His(Mtr)-OH After 3.4 g of Z-His(Mtr)-OH・DCHA was dissolved in 100 ml of ethyl acetate, 6 ml of 1N sulfuric acid was added, and then water was added and shaken. After mixing, the mixture was dried with anhydrous sodium sulfate. After distilling off the solvent, the mixture was dissolved in 100 ml of methanol for catalytic reduction. When the solvent was distilled off, crystals precipitated, so ether was added and the crystals were collected by filtration. Yield 1.80g (98.3%) Melting point 162-164℃ [α] 23 D -27.1゜ (C = 1.01, acetic acid), Rf 2 0.35 Elemental analysis Calculated as C 16 H 20 O 5 N 3 S・1/3H 2 O Value: C 51.60; H 5.55; N 11.28; S 8.61 Experimental value: C 51.76; H 5.63; N 11.08; S 8.63 Example 1 Bird GRP (Production of gastrin-releasing peptide (1) BOC-Leu-MetNH 2 Production of BOC-MetNH 2 (10.3g) with 4N-HCl/acetic acid 40
ml was added, and when the mixture was shaken, crystals precipitated, so ether was added, and the crystals were collected by filtration and dried.
Dissolve this in 200ml of DMF, cool on ice, and 7.0ml of TEA.
was added to neutralize it. In this, BOC−Leu−ONB
(BOC-Leu-OH8.0g, HONB6.85g,
(prepared from 7.83 g of DCC) was added and stirred overnight. After distilling off the solvent, dissolve in ethyl acetate,
After washing with aqueous sodium bicarbonate and citric acid, it was dried using anhydrous sodium sulfate. When the solvent was distilled off, crystals were precipitated, so ether was added, the crystals were collected by filtration, and recrystallized from methanol-ether. Yield 10.8g (86.3%) Melting point 152-154℃, [α] 23D -34.2゜(C=1.03,
DMF), Rf 1 0.65 Elemental analysis as C 16 H 31 O 4 N 3 S Calculated value: C 53.15; H 8.64; N 11.63; S 8.87 Experimental value: C 53.56; H 8.72; N 11.47; S 8.92 (2) BOC Production of His(Mtr)OH/DCHA 5.11 g of BOC-His OH was dissolved in a mixture of 30 ml of water and 30 ml of setone, and cooled on ice. Add this to 5.6ml of TEA
After adding Mtr-Cl4.97g acetone solution 30
ml and stirred for 2 hours. After acetone was distilled off under reduced pressure, the residue was acidified with citric acid, extracted with ethyl acetate, washed with water, and then dried using anhydrous sodium sulfate. After evaporating the solvent, it was dissolved in a small amount of ethyl acetate, 3.6 ml of DCHA was added, the solvent was evaporated, and the mixture was left overnight in a refrigerator. Ether was added to the precipitated crystals, which were collected by filtration. Yield 7.80g
(60.1%) Melting point 136-137°C, [α] 23 D +18.8° (c=1.01,
Methanol), Rf 1 0.63 Elemental analysis as C 33 H 52 O 7 N 4 S Calculated value: C 61.08; H 8.08; N 8.64; S 4.94 Experimental value: C 61.19; H 8.05; N 8.89; S 4.73 (3) BOC Production of -His(Mtr)-Leu-MetNH 2 To 10.0 g of BOC-Leu-MetNH 2 was added 30 ml of 4N-HCl/acetic acid, and the mixture was left to stand at room temperature for 20 minutes, then ether was added to form a precipitate, which was collected by filtration and dried. on the other hand
16.0 g of BOC-His(Mtr)OH・DCHA was suspended in ethyl acetate, and 27 ml of 1N sulfuric acid was added to separate the liquids.
It was dried using anhydrous sodium sulfate. After distilling off the solvent, dissolve in 100ml of acetonitrile,
After adding 5.0 g of HONB and cooling on ice, adding 5.8 g of DCC and reacting overnight, the precipitated DCU was filtered off. The previously prepared amine component was dissolved in 100 ml of DMF, neutralized with 4.6 ml of TEA, and then the active ester was added and stirred for 6 hours. After distilling off the solvent, it was dissolved in ethyl acetate, and dissolved in sodium bicarbonate solution, citric acid solution,
The mixture was washed successively with water and dried over anhydrous sodium sulfate. After distilling off the solvent, ether was added and the powder was collected by filtration. Yield 15.1g (86.0%) Melting point 129-131°C, [α] 23 D -13.8° (c=0.87,
DMF), Rf 1 0.64 Elemental analysis as C 32 H 50 O 8 N 6 S 2 Calculated value: C 54.06; H 7.09; N 11.82; S 9.02 Experimental value: C 54.69; H 7.39; N 11.97; S 8.39 (4) Production of Z-Val-Gly-OBu t After catalytic reduction of 12.0 g of Z-Gly-OBu t in methanol, the solvent was distilled off, and the residue was dissolved in DMF and cooled on ice.
To this, Z-Val-OH8.80g, HONB7.20g,
Added 8.24 g of DCC and stirred overnight. precipitated
After removing DCU by filtration, the residue was dissolved in ethyl acetate, washed with aqueous sodium bicarbonate and citric acid, and then dried over anhydrous sodium sulfate. The solvent was distilled off, petroleum benzine was added to the residue, the crystals were collected by filtration, and then recrystallized from ethyl acetate-petroleum benzine.
Yield 11.1g (87.0%) Melting point 141℃, [α] 23 D -2.1゜ (c=1.16, DMF),
Rf 1 0.80 Elemental analysis As C 19 H 28 O 5 N 2 Calculated value: C 62.62; H 7.74; N 7.69 Experimental value: C 62.49; H 7.60; N 7.72 (5) Z−Ala−Val−Gly−OBu t Production Z-Val-Gly-OBu t 10.0g in methanol,
After catalytic reduction, dissolve in 100ml of DMF, Z-
5.7 g of Ala-OH and 4.1 g of HONB were added and cooled on ice.
6.2g of DCC was added to this and stirred overnight. When the precipitated DCU was filtered off and the solvent was distilled off, crystals were precipitated, so ethyl acetate was added, the crystals were collected by filtration, and thoroughly washed. Yield 10.5g (96.4%) Melting point 184-185℃, [α] 23 D -7.8゜ (c=1.06,
DMF), Rf 1 0.67 Elemental analysis as C 22 H 33 O 6 N 3 Calculated value: C 60.67; H 7.64; N 9.65 Experimental value: C 60.93; H 7.86; N 9.77 (6) Z−Trp(Mtr)−Ala -Production of Val-Gly-OBu t After catalytic reduction of 5.0 g of Z-Ala-Val-Gly-OBu t in methanol, the solvent was distilled off and DMF100
It was dissolved into ml. In addition, Z-Trp (Mtr) OH6.34
g, 1.86 g of HOBT was added, cooled on ice, 2.85 g of DCC was added, and the mixture was stirred overnight. Filter off the precipitated DCU,
After distilling off the solvent, the extract was extracted with ethyl acetate containing a small amount of n-butanol, washed with aqueous sodium bicarbonate, and then dried over anhydrous sodium sulfate. The solvent was distilled off, ether was added, the precipitate was collected by filtration, and then crystals were collected from methanol-ethyl acetate-ether by filtration. Yield 8.80g (96.6%) Melting point 154-155℃, [α] 23 D -22.7゜ (c=0.88,
DMF), Rf 1 0.68 Elemental analysis as C 43 H 56 O 10 N 5 S Calculated value: C 61.85; H 6.76; N 8.39; S 3.84 Experimental value: C 62.06; H 7.01; N 8.58; S 3.58 (7) BOC −His(Mtr)−Trp(Mtr)−Ala−Val−
Production of Gly-OBu t Z-Trp (Mtr)-Ala-Val-GlyOBu t 8.0g
After catalytic reduction in DMF-methanol, methanol was distilled off. In this, BOC−His(Mtr)OH
(Prepared from 6.22g of BOC-His(Mtr)OH・DCHA), 1.90g of HONB was added and cooled on ice,
Added 2.18 g of DCC and stirred overnight. precipitated
After DCU was filtered off, the solvent was distilled off, ether was added, and a precipitate was collected by filtration. This was washed with a mixture of methanol, ethyl acetate, and ether.
Yield 10.2g (92.5%) Melting point 202-204℃, [α] 23 D -16.1゜ (c=1.18,
DMF), Rf 1 0.68 Elemental analysis as C 56 H 77 O 14 N 8 S 2 Calculated value: C 58.46; H 6.75; N 9.74; S 5.57 Experimental value: C 58.32; BOC−Ser−His(Mtr)−Trp(Mtr)−Ala−
Production of Val−GlyOH BOC−His(Mtr)−Trp(Mtr)−Ala−Val−
After adding 30 ml of TFA to 4.0 g of GlyOBu t and shaking at room temperature for 50 minutes, TFA was distilled off, ether was added, and the precipitate was collected by filtration and dried. On the other hand, BOC−
Ser−OH0.76g, HONB0.72g in acetonitrile
The mixture was dissolved to 20 ml, cooled on ice, and 0.83 g of DCC was added thereto, followed by stirring for 4 hours. Add the amine component prepared earlier to
The mixture was dissolved in 50 ml of DMF, neutralized by adding 1.0 ml of TEA, and then the active ester was added and stirred overnight. After distilling off the solvent, add a small amount of acetic acid,
Further water was added to form a precipitate which was collected by filtration. this
Re-precipitated from DMF-water. Yield 3.60g (85.0
%) Melting point 148-152℃, [α] 23 D -16.4゜(c=1.09,
DMF) Elemental analysis C 55 H 74 O 16 N 9 S 2・2H 2 O Calculated value: C 54.26; H 6.46; N 10.36; S 5.27 Experimental value: C 54.58; H 6.33; N 10.82; S 5.51 (9) Production of Z-Arg(Pme)-Gly-OBu t After catalytic reduction of 13 g of Z-Gly・OBu t in 300 ml of methanol, the solvent was distilled off and the residue was
Dissolved in 200ml of DMF. To this, Z−Arg(Pme)
OH [Prepared from 20g of Z-Arg(Pme)OH・CHA]
was added, cooled on ice, 5.4 g of HOBT and 8.2 g of DCC were added, and the mixture was stirred overnight. After the precipitated DCU was filtered off, the solvent was distilled off, and the residue was dissolved in ethyl acetate. This was washed with aqueous sodium bicarbonate and citric acid, and then dried over anhydrous sodium sulfate. After distilling off the solvent, petroleum benzine was added and the powder was collected by filtration. Yield 19.8g (95.0%) Melting point 55-60℃, [α] 23 D +0.2゜ (c=0.88,
DMF), Rf 1 0.62 Elemental analysis as C 31 H 45 O 7 N 5 S Calculated value: C 58.93; H 7.18; N 11.09; S 5.08 Experimental value: C 58.96; H 7.01; N 10.67; S 5.05 (10) Z Production of -Tyr-Pro-OBu t 15.0 g of Z-Pro OBu t was dissolved in 300 ml of methanol for catalytic reduction, the solvent was distilled off, and 400 ml of DMF was added.
I grinned. To this, Z-Tyr-OH (Z-Tyr-
(prepared from 20.0 g of OH/DCHA) and 6.75 g of HOBT were added and cooled on ice. Further, 10.4 g of DCC was added and stirred overnight. After the precipitated DCU was filtered off and the solvent was distilled off, it was dissolved in ethyl acetate, washed with aqueous sodium bicarbonate and citric acid, and dried over anhydrous sodium sulfate. After distilling off the solvent, it was applied to a silica gel column (7.5 x 9 cm) and eluted with 1% (MeOH/chloroform). Fractions of the target product were collected and concentrated, and then petroleum benzine was added and the powder was collected by filtration. did.
Yield 15.4g (82.2%) [α] 23 D −39.9° (c=0.83, DMF) Rf 1 0.62 Elemental analysis C 26 H 32 O 6 N 2・1/2H 2 O Calculated value: C 65.39; H 6.97 ; N 5.87 Experimental value: C 65.70; H 6.93; N 5.66 (11) Z-Ile-Tyr-Pro-OBu t Z-Tyr-Pro-OBu t 15.2g in methanol 300
ml, and after catalytic reduction, the solvent was distilled off,
The residue was dissolved in 200ml DMF. Z- to this
After adding 8.0 g of IleOH and 6.5 g of HONB and cooling on ice,
Added 7.4 g of DCC and stirred overnight. a small amount of N,
After adding N-dimethylpropanediamine,
DCU was filtered off and the solvent was distilled off under reduced pressure. The residue was dissolved in ethyl acetate, washed with aqueous sodium bicarbonate and citric acid, and dried over anhydrous sodium sulfate. When the solvent is distilled off, crystals will precipitate, so petroleum benzine is added and the crystals are collected by filtration, followed by methanol and
Recrystallized from ether and petroleum benzine. yield
10.9g (62.5%) Melting point 177-178℃, [α] 23 D -38.3゜(c=1.11,
DMF), Rf 1 0.62 Elemental analysis as C 32 H 43 O 7 N 3 Calculated value: C 66.07; H 7.45; N 7.22 Experimental value: C 66.07; H 7.74; N 7.19 (12) Z−Ile−Try−Pro− Production of OH 60 ml of TFA was added to 6.0 g of Z-Ile-Tyr-Pro OBu t and stirred at room temperature for 1 hour, then distilled off. The residue was dissolved in ethyl acetate, washed with water, and then dried over anhydrous sodium sulfate. After distilling off the solvent, ether was added to form a powder, which was collected by filtration. Yield 5.10g (94.2%) Melting point 72-74℃, [α] 23 D -25.2゜ (c=1.01,
DMF), Rf 1 0.44 Elemental analysis as C 28 H 35 O 7 N 3 Calculated value: C 63.98; H 6.71; N 7.99 Experimental value: C 63.75; H 6.67; N 7.84 (13) Z−Ile−Tyr−Pro− Arg(Pme)−
Production of GlyOBu t Dissolve 7.58 g of Z-Arg(Pme)-Gly-OBu t in 300 ml of methanol, add 12 ml of 1N-hydrochloric acid,
Contact reduction. After distilling off the solvent, remove the residue.
The mixture was dissolved in 100 ml of DMF, cooled on ice, and neutralized by adding 2.00 ml of TEA. To this, Z-Ile-Tyr-Pro-
Add OH5.78g, HOBT2.23g, DCC3.40g,
I stirred it all night. After filtering off the precipitated DCU,
The solvent was evaporated and the residue was dissolved in ethyl acetate.
This was washed with aqueous sodium bicarbonate and citric acid, and dried over anhydrous sodium sulfate. The solvent was distilled off, the residue was applied to a silica gel column (5.5 x 10 cm), and 2%
After elution with MeOH/chloroform, fractions of the target product were collected and concentrated, ether was added, and the powder was collected by filtration. Yield 7.20g (62.2%) Melting point 110-112℃, [α] 23 D -28.3゜(c=1.19,
DMF), Rf 1 0.59 Elemental analysis C 51 H 72 O 11 N 8 S・H 2 O Calculated value: C 59.85; H 7.29; N 10.93; S
3.13 Experimental value: C 60.16; H 7.56; N 10.95; S
2.98 (14) Z−Lys(Mtr)−Ile−Tyr−Pro−Arg
Production of (Pme)−GlyOBu t Z−Ile−Tyr−Pro−Arg(Pme)−Gly−OBu t
Dissolve 7.0g in 350ml of methanol and add 7.0g of 1N-hydrochloric acid.
ml was added for catalytic reduction. After distilling off the solvent, dissolve the residue in 200 ml of DMF, cool on ice,
1.0 ml of TEA was added to neutralize. To this, Z-Lys
(Mtr)-OH [Z-Lys(Mtr)OH・DCHA4.74g
], add 120 g of HOBT, 1.80 g of DCC,
I stirred it all night. The precipitated DCU was filtered off and the residue was dissolved in ethyl acetate. This was washed with aqueous sodium bicarbonate and citric acid, and then dried over anhydrous sodium sulfate. The solvent was distilled off, ether was added to form a powder, and the powder was reprecipitated twice from acetic ether-ether. Yield 8.40g (87.5%) Melting point 116-118℃, [α] 23 D -19.9゜ (c=1.05,
DMF), Rf 1 0.59 Elemental analysis as C 66 H 96 O 15 N 10 S 2 Calculated value: C 59.44; H 7.26; N 10.50; S 4.81 Experimental value: C 59.10; H 7.44; N 10.46; S 4.98 (15) Z−Thr−Lys(Mtr)−Ile−Tyr−Pro−
Production of Arg(Pme)−Gly−OBu t Z−Lys(Mtr)−Ile−Tyr−Pro−Arg(Pme)
- After catalytic reduction of 7.50 g of GlyOBu t in 350 ml of methanol, the solvent was distilled off and the residue was dissolved in 100 ml of DMF. To this, Z-Thr-OH1.49g,
After adding HONB1.51g and cooling on ice, DCC1.73
g and stirred overnight. After adding a small amount of N,N-dimethylpropanediamine and filtering off DCU, the solvent was distilled off and the residue was dissolved in ethyl acetate containing a small amount of n-butanol. This was washed with aqueous sodium bicarbonate, then water, and dried over anhydrous sodium sulfate. The solvent was distilled off, and ether was added to the residue to form a powder, which was then collected by filtration. yield
8.0g (99.2%) Melting point 122-124℃, [α] 23 D -25.5゜(c=0.82,
DMF), Rf 1 0.59 Elemental analysis C 70 H 103 O 17 N 11 S 2 Calculated value: C 58.60; H 7.24; N 10.74; S
4.47 Experimental value: C 58.61; H 7.29; N 10.47; S
4.02 (16) BOC−Leu−Thr−Lys(Mtr)−Ile−Tyr−
Production of Pro−Arg(Pme)−GlyOBu t Z−Thr−Lys(Mtr)−Ile−Tyr−Pro−Arg
7.5 g of (Pme)-GlyOBu t was dissolved in 350 ml of methanol, and 0.99 g of para-toluenesulfonic acid was added for catalytic reduction, and then the solvent was distilled off. this
The mixture was dissolved in 100 ml of DMF, cooled on ice, and neutralized by adding 0.74 ml of TEA. To this, 1.37g of BOC-Leu-OH,
1.41 g of HONB and 1.62 g of DCC were added and stirred overnight. After DCU was filtered off and the solvent was distilled off, it was dissolved in ethyl acetate, washed with aqueous sodium bicarbonate and citric acid, and dried over anhydrous sodium sulfate. After distilling off the solvent, ether was added and the powder was collected by filtration. Yield 7.55g (95.4%) Melting point 136-138℃, [α] 23 D -26.6゜ (c=1.00,
DMF), Rf 1 0.59 Elemental analysis C 73 H 116 O 18 N 12 S 2・H 2 O Calculated value: C 57.23; H 7.76; N 10.97; S 4.19 Experimental value: C 57.53; H 8.12; N 10.65; S 3.90 (17) BOC−Ala−Leu−Thr−Lys(Mtr)−Ile−
Production of Tyr−Pro−Arg(Pme)−Gly−OH BOC−Leu−Thr−Lys(Mtr)−Ile−Tyr−Pro
Add 40 ml of TFA to 4.20 g of -Arg(Pme)-Gly-OBu t , stir at room temperature for 60 minutes, and then evaporate the solvent.
Ether was added to the powder, which was collected by filtration and dried.
Dissolve this in 50ml of DMF, cool on ice, and 0.80ml of TEA.
added. In addition, BOC−Ala−ONB (BOC−
(Prepared from 0.52 g of Ala-OH, 0.54 g of HONB, and 0.62 g of DCC) was added and stirred overnight. a small amount of N,
After adding N-dimethylpropanediamine and distilling off the solvent, aqueous acetic acid was added to form a precipitate, which was collected by filtration. This was reprecipitated with aqueous methanol. yield
3.60g (88.1%) Melting point 128-130℃, [α] 23 D -32.9゜(c=0.98,
DMF), Rf 1 0.40 Elemental analysis C 72 H 113 O 19 N 13 S 2・2H 2 O Calculated value: C 55.26; H 7.54; N 11.64; S 4.10 Experimental value: C 55.02; H 6.89; N 11.38; S 3.77 (18) Production of Z-Ser-ProOBu t 11.0 g of Z-Pro-OBu t was dissolved in 300 ml of methanol, and after catalytic reduction, the solvent was distilled off,
Dissolved in 200ml of DMF. Z-Ser-OH7.2 to this
g, 4.90 g of HOBT, and 7.50 g of DCC were added and stirred overnight. The precipitated DCU was filtered off, the solvent was distilled off, and the residue was dissolved in ethyl acetate. This was washed with aqueous sodium bicarbonate and citric acid, and then dried over anhydrous sodium sulfate. The solvent was distilled off, petroleum benzine was added, the crystals were collected by filtration, and then recrystallized from ethyl acetate-ether. Yield 9.50g (67.2%) Melting point 126-127℃, [α] 23 D -50.9゜(c=0.95,
DMF), Rf 1 0.65 Elemental analysis as C 20 H 28 O 6 N 2 Calculated value: C 61.21; H 7.19; N 7.14 Experimental value: C 61.45; H 7.16; N 7.31 (19) Z−Gly−Ser−Pro− Production of OBu t 10.0g of Z-Ser-Pro-OBu t in 300ml of methanol
After stirring and catalytic reduction, the solvent was distilled off.
Dissolved in 300ml of DMF. Z-Gly-OH5.06 for this
g、After adding 5.13 g of HONB and cooling on ice,
Added 5.89 g of DCC and stirred overnight. After adding a small amount of N,N-dimethylpropanediamine, DCU was filtered off and the solvent was distilled off. The residue was dissolved in ethyl acetate, washed with aqueous sodium bicarbonate and citric acid, and dried over anhydrous sodium sulfate. The solvent was distilled off, petroleum benzine was added, the crystals were collected by filtration, and then recrystallized from ether. Yield 7.70g (72.1
%) Melting point 96-98℃, [α] 23 D -53.4゜(c=1.05,
DMF), Rf 1 0.61 Elemental analysis as C 22 H 31 O 7 N 3 Calculated value: C 58.78; H 6.95; N 9.35 Experimental value: C 58.86; H 7.04; N 9.46 (20) Z−Gly−Gly−Ser− Production of Pro-OBu t 7.0g of Z-Gly-Ser-Pro-OBu t was added to methanol.
After dilution to 200 ml and catalytic reduction, the solvent was distilled off and the residue was dissolved in 100 ml of DMF. Z- to this
After adding 3.0 g of Gly-OH and 3.10 g of HONB and cooling on ice, 3.60 g of DCC was added and stirred overnight. After adding a small amount of N,N-dimethylpropanediamine, DCU was filtered off and the solvent was distilled off. The residue was dissolved in ethyl acetate, washed with saturated brine, and then dried over anhydrous sodium sulfate. The solvent was distilled off and ether was added to form crystals, which were collected and recrystallized from ethyl acetate. Yield 5.70g (78.5%) Melting point 129-130℃, [α] 23 D -47.0゜ (c=0.81,
DMF), Rf 1 0.47 Elemental analysis as C 24 H 34 O 8 N 4 Calculated value: C 56.90; H 6.77; N 11.06 Experimental value: C 56.75; H 6.68; N 10.90 (21) Z−Gln−Pro−OBu t Production of Z-Pro-OBu t 16.2g was dissolved in 350ml of methanol, and after catalytic reduction, the solvent was distilled off and DMF200
It was dissolved into ml. To this, Z-Gln-OH12.4g,
7.16 g of HOBT was added, cooled on ice, 10.9 g of DCC was added, and the mixture was stirred overnight. The precipitated DCU was filtered off,
After evaporating the solvent, it was dissolved in ethyl acetate, washed with aqueous sodium bicarbonate and citric acid, and dried over anhydrous sodium sulfate. After distilling off the solvent, petroleum benzine was added to form crystals, which were collected by filtration and recrystallized from ethyl acetate-petroleum benzine. Yield 15.8g (82.5%) Melting point 106-107℃, [α] 23 D -51.2゜ (c=1.06,
DMF), Rf 1 0.62 Elemental analysis as C 22 H 31 O 6 N 3 Calculated value: C 60.95; H 7.21; N 9.69 Experimental value: C 60.95; H 7.36; N 9.41 (22) Z−Leu−Gln−Pro− Production of OBut 8.0 g of Z-Gln-Pro-OBu t and 3.51 g of para-toluenesulfonic acid were dissolved in 300 ml of methanol for catalytic reduction, and then the solvent was distilled off. DMF200 residue
ml, cooled on ice, and neutralized by adding 2.6 ml of TEA. To this, Z-LeuOH (Z-Leu-OH・
Prepared from DCHA8.24g), HONB4.00g,
Added 4.60 g of DCC and stirred overnight. a small amount of N,
After adding N-dimethylpropanediamine
DCU was filtered off and the solvent was distilled off. Dissolve the residue in ethyl acetate, wash with sodium bicarbonate solution and citric acid solution,
It was dried with anhydrous sodium sulfate. After the solvent was distilled off to precipitate crystals, petroleum benzine was added and the crystals were collected by filtration and recrystallized from ethyl acetate-petroleum benzine. Yield 7.70g (76.3%) Melting point 62-64℃, [α] 23 D -51.7゜ (c=1.11,
DMF), Rf 1 0.61 Elemental analysis as C 28 H 42 O 7 N 4 Calculated value: C 61.52; H 7.75; N 10.25 Experimental value: C 61.19; H 7.75; N 10.11 (23) BOC−Pro−Leu−Gln− Production of Pro-OBu t 7.0 g of Z-Leu-Gln-Pro-OBu t was dissolved in 300 ml of methanol and subjected to catalytic reduction, the solvent was distilled off, and the residue was dissolved in 100 ml of DMF. to this
After adding 2.42 g of BOC-ProOH and 2.76 g of HONB and cooling on ice, 3.17 g of DCC was added and stirred overnight. After adding a small amount of N,N-dimethylpropanediamine, the DCU was filtered off, the solvent was evaporated and dissolved in ethyl acetate. This was washed with aqueous sodium bicarbonate and citric acid, and then dried over anhydrous sodium sulfate. After distilling off the solvent, the powder was filtered from ether-petroleum benzine. Yield 6.50g (83.3
%) Melting point 74-76℃, [α] 23 D -76.3゜(c=1.19,
DMF), Rf 1 0.60 Elemental analysis C 30 H 51 O 8 N 5 Calculated value: C 59.05; H 8.43; N 11.49 Experimental value: C 58.89; H 8.12; N 11.08 (24) BOC−Ala−Pro−Leu− Production of Gln-Pro-OH BOC-Pro-Leu-Gln-Pro-OBu t 6.0g
After adding 60ml of TFA and stirring at room temperature for 60 minutes,
The solvent was distilled off, and ether was added to the residue to form a powder, which was filtered and dried. Dissolve this in 100ml of DMF, cool on ice, add 2.80ml of TEA, and add BOC-Ala.
-ONB (BOC-Ala-OH1.95g, HONB2.07g,
(prepared from 2.38 g of DCC) was added and stirred overnight.
After distilling off the solvent, a small amount of acetic acid was added, followed by ether, and the mixture was filtered as a powder. Dissolve this in chloroform and silica gel column (5.5
x 8 cm), eluted with 5% methanol/chloroform, collected a fraction of the target substance, and concentrated.
Ether was added and the powder was collected by filtration. Yield 4.50
g (73.2%) Melting point 124-128℃, [α] 23 D -82.1゜(c=1.04,
DMF), Rf 1 0.18 Elemental analysis as C 29 H 48 O 9 N 6 Calculated value: C 55.75; H 7.74; N 13.45 Experimental value: C 55.38; H 7.65; N 13.21 (25) BOC−Ala−Pro−Leu− Gln−Pro−Gly−
Production of Gly-Ser-Pro-OBu t 2.23 g of Z-Gly-Gly-Ser-Pro-OBu t was dissolved in 100 ml of methanol for catalytic reduction, the solvent was distilled off, and the residue was dissolved in 50 ml of DMF. to this
BOC-Ala-Pro-Leu-Gln-Pro-OH2.50g,
Add 0.90g of HONB, cool on ice, and add 1.03g of DCC.
and stirred overnight. After filtering off the precipitated DCU, the solvent was distilled off and a small amount of n-BuOH was added.
The mixture was dissolved in ethyl acetate containing 100% chloride, washed with saturated brine, and dried over anhydrous sodium sulfate. After distilling off the solvent, ether was added and the powder was collected by filtration. Yield 3.85g (98.3%) Melting point 100-105℃, [α] 23 D -74.0゜ (c=0.92,
DMF), Rf 1 0.19 Elemental analysis as C 45 H 74 O 14 N 10・H 2 O Calculated value: C 54.20; H 7.68; N 14.05 Experimental value: C 54.01; H 7.45; N 13.44 (26) BOC−Ala− Pro−Leu−Gln−Pro−Gly−
Production of Gly−Ser−Pro−OH BOC−Ala−Pro−Leu−Gln−Pro−Gly−Gly
10 ml of TFA was added to 1.0 g of -Ser-Pro-OBu t and the mixture was stirred at room temperature for 1 hour. The solvent was distilled off, ether was added, and the mixture was filtered as a powder. Add this to DMF10
ml, cool on ice, then add 0.46ml of TEA.
Next, 0.27 g of BOC-OH was added and stirred for 4 hours. After distilling off the solvent, 1.5 ml of acetic acid was added, and ether was added to obtain a powder, which was collected by filtration. This was reprecipitated from methanol-ether. Yield 0.87g
(92.4%) Melting point 141-145°C, [α] 23 D -72.9° (c=0.98,
DMF), Rf 3 0.29 Elemental analysis C 41 H 66 O 14 N 10・3H 2 O Calculated value: C 50.40; H 7.43; N 14.34 Experimental value: C 50.16; H 6.76; N 14.11 (27) BOC−Ser− His(Mtr)−Trp(Mtr)−Ala
-Production of Val-Gly-His(Mtr)-Leu-Met- NH2 BOC-His(Mtr)-Leu- MetNH2 3.97g
After adding 40ml of TFA and stirring at room temperature for 10 minutes,
4.1 ml of 1.3N hydrochloric acid was added, the solvent was distilled off, ether was added, the powder was collected by filtration, and the mixture was dried. this
Dissolve in 40ml of DMF, cool on ice, add 0.86ml of TEA, then BOC-Ser-His(Mtr)-Trp(Mtr)-
Ala-Val-Gly-OH5.50g, HONB1.0g,
Added 1.15 g of DCC and stirred overnight. precipitated
After DCU was filtered off and the solvent was distilled off, water was added and the powder was collected by filtration. This was then washed with aqueous ethal. Yield 7.35g (88.9%) Melting point 192-193℃, [α] 23 D -12.5゜ (c=1.05,
DMF), Rf 1 0.50 Elemental analysis C 82 H 114 O 21 N 15 S 4 Calculated value: C 55.51; H 6.48; N 11.84; S 7.23 Experimental value: C 55.56; H 6.74; N 11.83; S 6.55 (28) BOC−Ser−His−Trp(Mtr)−Ala−Val
−Production of Gly−His−Leu−Met−NH 2 BOC−Ser−His(Mtr)−Trp(Mtr)−Ala−
Val−Gly−His(Mtr)−Leu−Met−NH 2 3.0g
The mixture was dissolved in 15 ml of DMF, 2.30 g of HOBT was added, and the mixture was treated for 30 minutes. The solvent was distilled off, ether was added, and a powder was collected by filtration. Yield 2.15g (92.5%) Melting point 191-193℃, [α] 23 D -18.8゜ (c=0.94,
DMF), Rf 3 0.64 Elemental analysis C 62 H 90 O 15 N 15 S 2・H 2 O Calculated value: C 54.45; H 6.78; N 15.36; S 4.69 Experimental value: C 53.92; H 6.43; N 15.80; S 4.34 (29) BOC−Ala−Leu−Thr−Lys(Mtr)−Ile−
Tyr−Pro−Arg(Pme)−Gly−Ser−His−
Trp(Mtr)−Ala−Val−Gly−His−Leu−Met
−Production of NH 2 BOC−Ser−His−Trp(Mtr)−Ala−Val−
20 ml of TFA was added to 2.0 g of Gly-His-Leu-MetNH 2 and treated at room temperature for 15 minutes, then distilled off, ether was added, the powder was collected by filtration, and dried. this
The mixture was dissolved in 5 ml of DMF, 1.24 ml of TEA was added, and the mixture was stirred well. Ether was added to precipitate the mixture, which was filtered as a powder. Dissolve this in 20ml of ether,
BOC−Ala−Leu−Thr−Lys(Mtr)−Ile−Tyr−
Pro−Arg(Pme)−Gly−OH1.78g, HONB0.41
After adding g, the mixture was cooled on ice, 0.47 g of DCC was added, and the mixture was stirred overnight. After distilling off the solvent, ethanol-
After adding ethyl acetate and filtering the powder, it was washed with hot ethanol. Yield 3.30g (99.7%) Melting point 222-223℃ (decomposition), [α] 23D -22.8゜(c
=
1.02, DMF), Rf 3 0.67 Elemental analysis C 129 H 193 O 31 N 28 S 4・8H 2 O Calculated value: C 53.34; H 7.25; N 13.50; S 4.42 Experimental value: C 53.35; H 6.98; N 12.91 ;S 4.58 (30) BOC−Ala−Pro−Leu−Gln−Pro−Gly−
Gly−Ser−Pro−Ala−Leu−Thr−Lys(Mtr)
−Ile−Tyr−Pro−Arg(Pme)Gly−Ser−His
(Mtr)−Trp(Mtr)−Ala−Val−Gly−His−
Production of Leu−Met−NH 2 BOC−Ala−Leu−Thr−Lys(Mtr)−Ile−Tyr
−Pro−Arg(Pme)−Gly−Ser−His−Trp
(Mtr) −Ala−Val−Gly−His−Leu−Met−
Add 5ml of TFA to 500mg of NH 2 , mix well,
The residue was distilled off, ether was added, and the powder was collected by filtration.
Dissolve this in 1ml of DMF, add 0.2ml of TEA,
After stirring well, ether was added and the mixture was filtered as a powder. Dissolve this in 5ml of DMF and add BOC.
−Ala−Pro−Leu−Gln−Pro−Gly−Gly−Ser
After adding 217 mg of -Pro-OH and 50 mg of HOBT, the mixture was cooled on ice, and 160 mg of DCC was added and stirred overnight. After the precipitated DCU was filtered off, the solvent was distilled off, and ethyl acetate was added to the residue, which was filtered as a powder. After dissolving this in DMF-methanol and heating for about 30 minutes, methanol was distilled off, ethyl acetate was added to the residue, and the powder was collected by filtration. This was then washed with methanol water. Yield 480mg (72.2%) Melting point 203-208℃ (decomposition), [α] 23D -35.2゜(c
=
0.93, DMF), Rf 3 0.66 Elemental analysis C 165 H 249 O 42 N 38 S 4・6H 2 O Calculated value: C 53.95; H 7.16; N 14.49; S 3.49 Experimental value: C 54.08; H 6.87; N 14.19 ;S 3,43 (31) H-Ala-Pro-Leu-Gln-Pro-Gly-
Gly−Ser−Pro−Ala−Leu−Thr−Lys−Ile
−Tyr−Pro−Arg−Gly−Ser−His−Trp−
Ala−Val−Gly−His−Leu−Met−NH 2 (bird
BOC−Ala−Pro−Leu−Gln−Pro−Gly−Gly
−Ser−Pro−Ala−Leu−Thr−Lys(Mtr)−Ile
−Tyr−Pro−Arg(Pme)−Gly−Ser−His−
Trp (Mtr) −Ala−Val−Gly−His−Leu−Met−
Add 8 ml of 0.3 M methanesulfonic acid/TFA-thioanisole (9:1) to 100 mg of NH 2 , stir at room temperature for 2 hours, add 100 mg of ammonia acetate, distill off TFA, and add ether to form a powder. I took it as a toro. Dissolve this in a small amount of 1N acetic acid water,
It was applied to a Sephadex G-25 column (2.2 x 120 cm). Elute with 1N acetic acid and take a fraction of 170ml to 270ml, freeze-dry, and dissolve in a small amount of water.
After passing through an Amberlite IRA410 column (1 x 10 cm), it was applied to a carboxymethyl cellulose column (2.2 x 17 cm). Elute with a linear gradient of water (400 ml) - 0.4 M ammonia acetate (400 ml).
Fractions of 335-365 ml were collected and lyophilized. Yield 28
mg (35%). [α] 22 D -102.2゜(c=0.32, 1% acetic acid) Rf 4 0.39 Amino acid analysis value (4% thioglycolic acid/6N
Hydrochloric acid hydrolysis): Lys1.00(1); His1.73(2); Arg1.04
(1);Trp0.67(1);Thr1.06(1);Ser1.77(2);Glu1.11
(1);Pro4.20(4);Gly4.23(4);Ala3.19(3);Val1.06
(1);Met1.02(1);Ile0.96(1);Leu3.04(3);Tyr1.00
(1) (Average response rate 73.7%)
Claims (1)
り、ヒスチジンのイミダゾール基を、４−メトキ
シ−２，３，６−トリメチルベンゼンスルホニル
基で保護し、ペプチド縮合したのち、該保護基を
酸または１−ヒドロキシベンゾトリアゾールで脱
離せしめることを特徴とするペプチドの製造法。 ２ 酸としてトリフルオロ酢酸を用いる特許請求
の範囲第１項記載の製造法。 ３ チオアニソールまたはジメチルスルフイドの
存在下で保護器を脱離せしめる特許請求の範囲第
１項または第２項記載の製造法。[Claims] 1. In producing a peptide containing histidine, the imidazole group of histidine is protected with a 4-methoxy-2,3,6-trimethylbenzenesulfonyl group, and after peptide condensation, the protecting group is removed with an acid or 1. A method for producing a peptide, characterized by elimination with 1-hydroxybenzotriazole. 2. The manufacturing method according to claim 1, wherein trifluoroacetic acid is used as the acid. 3. The manufacturing method according to claim 1 or 2, wherein the protector is removed in the presence of thioanisole or dimethyl sulfide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1086538A JPH0249799A (en) | 1989-04-05 | 1989-04-05 | Production of peptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1086538A JPH0249799A (en) | 1989-04-05 | 1989-04-05 | Production of peptide |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP56174124A Division JPS5874650A (en) | 1981-10-29 | 1981-10-29 | Preparation of peptide and histidine derivative |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0249799A JPH0249799A (en) | 1990-02-20 |
| JPH0224836B2 true JPH0224836B2 (en) | 1990-05-30 |
Family
ID=13889778
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1086538A Granted JPH0249799A (en) | 1989-04-05 | 1989-04-05 | Production of peptide |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0249799A (en) |
-
1989
- 1989-04-05 JP JP1086538A patent/JPH0249799A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0249799A (en) | 1990-02-20 |
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