JPH07116228B2 - Process for producing α-L-aspartyl-L-phenylalanine methyl ester - Google Patents
Process for producing α-L-aspartyl-L-phenylalanine methyl esterInfo
- Publication number
- JPH07116228B2 JPH07116228B2 JP61305329A JP30532986A JPH07116228B2 JP H07116228 B2 JPH07116228 B2 JP H07116228B2 JP 61305329 A JP61305329 A JP 61305329A JP 30532986 A JP30532986 A JP 30532986A JP H07116228 B2 JPH07116228 B2 JP H07116228B2
- Authority
- JP
- Japan
- Prior art keywords
- aspartyl
- phenylalanine
- apm
- methanol
- formyl
- 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
- 238000000034 method Methods 0.000 title claims description 45
- IAOZJIPTCAWIRG-QWRGUYRKSA-N aspartame Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 IAOZJIPTCAWIRG-QWRGUYRKSA-N 0.000 title claims description 12
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 84
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 84
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 54
- YZQCXOFQZKCETR-UWVGGRQHSA-N Asp-Phe Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 YZQCXOFQZKCETR-UWVGGRQHSA-N 0.000 claims description 41
- 238000004519 manufacturing process Methods 0.000 claims description 30
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 26
- 238000005886 esterification reaction Methods 0.000 claims description 19
- WYYUBJAMROQJSF-QWRGUYRKSA-N (3s)-4-[[(1s)-1-carboxy-2-phenylethyl]amino]-3-formamido-4-oxobutanoic acid Chemical compound OC(=O)C[C@H](NC=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 WYYUBJAMROQJSF-QWRGUYRKSA-N 0.000 claims description 11
- VSDUZFOSJDMAFZ-VIFPVBQESA-N methyl L-phenylalaninate Chemical compound COC(=O)[C@@H](N)CC1=CC=CC=C1 VSDUZFOSJDMAFZ-VIFPVBQESA-N 0.000 claims description 10
- 238000011065 in-situ storage Methods 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 4
- 230000003472 neutralizing effect Effects 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 3
- 229960002337 magnesium chloride Drugs 0.000 description 24
- 238000006243 chemical reaction Methods 0.000 description 21
- 239000002609 medium Substances 0.000 description 21
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 13
- 239000002994 raw material Substances 0.000 description 11
- 230000000052 comparative effect Effects 0.000 description 10
- 229960005190 phenylalanine Drugs 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- DFTMVZIUYVECNW-VKHMYHEASA-N n-[(3s)-2,5-dioxooxolan-3-yl]formamide Chemical class O=CN[C@H]1CC(=O)OC1=O DFTMVZIUYVECNW-VKHMYHEASA-N 0.000 description 7
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 description 6
- 238000004128 high performance liquid chromatography Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 230000032050 esterification Effects 0.000 description 5
- IXCSERBJSXMMFS-UHFFFAOYSA-N hydrogen chloride Substances Cl.Cl IXCSERBJSXMMFS-UHFFFAOYSA-N 0.000 description 4
- 229910000041 hydrogen chloride Inorganic materials 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000035484 reaction time Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 3
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000009833 condensation Methods 0.000 description 3
- 230000005494 condensation Effects 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 229960005261 aspartic acid Drugs 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 229940050906 magnesium chloride hexahydrate Drugs 0.000 description 2
- DHRRIBDTHFBPNG-UHFFFAOYSA-L magnesium dichloride hexahydrate Chemical compound O.O.O.O.O.O.[Mg+2].[Cl-].[Cl-] DHRRIBDTHFBPNG-UHFFFAOYSA-L 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- MTQBJQWJDGTPIL-QWRGUYRKSA-N (2s)-4-[[(1s)-1-carboxy-2-phenylethyl]amino]-2-formamido-4-oxobutanoic acid Chemical compound O=CN[C@H](C(=O)O)CC(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MTQBJQWJDGTPIL-QWRGUYRKSA-N 0.000 description 1
- GWKOSRIHVSBBIA-REOHCLBHSA-N (3s)-3-aminooxolane-2,5-dione Chemical compound N[C@H]1CC(=O)OC1=O GWKOSRIHVSBBIA-REOHCLBHSA-N 0.000 description 1
- QDGAVODICPCDMU-UHFFFAOYSA-N 2-amino-3-[3-[bis(2-chloroethyl)amino]phenyl]propanoic acid Chemical compound OC(=O)C(N)CC1=CC=CC(N(CCCl)CCCl)=C1 QDGAVODICPCDMU-UHFFFAOYSA-N 0.000 description 1
- QCQCHGYLTSGIGX-GHXANHINSA-N 4-[[(3ar,5ar,5br,7ar,9s,11ar,11br,13as)-5a,5b,8,8,11a-pentamethyl-3a-[(5-methylpyridine-3-carbonyl)amino]-2-oxo-1-propan-2-yl-4,5,6,7,7a,9,10,11,11b,12,13,13a-dodecahydro-3h-cyclopenta[a]chrysen-9-yl]oxy]-2,2-dimethyl-4-oxobutanoic acid Chemical compound N([C@@]12CC[C@@]3(C)[C@]4(C)CC[C@H]5C(C)(C)[C@@H](OC(=O)CC(C)(C)C(O)=O)CC[C@]5(C)[C@H]4CC[C@@H]3C1=C(C(C2)=O)C(C)C)C(=O)C1=CN=CC(C)=C1 QCQCHGYLTSGIGX-GHXANHINSA-N 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- CKLJMWTZIZZHCS-UHFFFAOYSA-N D-OH-Asp Natural products OC(=O)C(N)CC(O)=O CKLJMWTZIZZHCS-UHFFFAOYSA-N 0.000 description 1
- CKLJMWTZIZZHCS-UWTATZPHSA-N L-Aspartic acid Natural products OC(=O)[C@H](N)CC(O)=O CKLJMWTZIZZHCS-UWTATZPHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- MQUUQXIFCBBFDP-VKHMYHEASA-N N-formyl-L-aspartic acid Chemical compound OC(=O)C[C@@H](C(O)=O)NC=O MQUUQXIFCBBFDP-VKHMYHEASA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229910001854 alkali hydroxide Inorganic materials 0.000 description 1
- 150000008044 alkali metal hydroxides Chemical class 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 239000008122 artificial sweetener Substances 0.000 description 1
- 235000021311 artificial sweeteners Nutrition 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 125000002843 carboxylic acid group Chemical group 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- -1 ester compound Chemical class 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 150000003840 hydrochlorides Chemical class 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- OJURWUUOVGOHJZ-UHFFFAOYSA-N methyl 2-[(2-acetyloxyphenyl)methyl-[2-[(2-acetyloxyphenyl)methyl-(2-methoxy-2-oxoethyl)amino]ethyl]amino]acetate Chemical compound C=1C=CC=C(OC(C)=O)C=1CN(CC(=O)OC)CCN(CC(=O)OC)CC1=CC=CC=C1OC(C)=O OJURWUUOVGOHJZ-UHFFFAOYSA-N 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 230000007030 peptide scission Effects 0.000 description 1
- BXRNXXXXHLBUKK-UHFFFAOYSA-N piperazine-2,5-dione Chemical class O=C1CNC(=O)CN1 BXRNXXXXHLBUKK-UHFFFAOYSA-N 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- REFMEZARFCPESH-UHFFFAOYSA-M sodium;heptane-1-sulfonate Chemical compound [Na+].CCCCCCCS([O-])(=O)=O REFMEZARFCPESH-UHFFFAOYSA-M 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000000967 suction filtration Methods 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts
Landscapes
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Low-Molecular Organic Synthesis Reactions Using Catalysts (AREA)
- Peptides Or Proteins (AREA)
- Catalysts (AREA)
Description
【発明の詳細な説明】 （産業上の利用分野） 本発明は、α−Ｌ−アスパルチル−Ｌ−フェニルアラニ
ンメチルエステル（以下、α−APMと略記する）の改良
された製造法に関する。TECHNICAL FIELD The present invention relates to an improved method for producing α-L-aspartyl-L-phenylalanine methyl ester (hereinafter abbreviated as α-APM).
さらに詳しくは、α−Ｌ−アスパルチル−Ｌ−フェニル
アラニンを塩酸とメタノールから成る媒体系でエステル
化し、生成した固体状のα−APM塩酸塩を固液分離し、
その後、この塩酸塩を中和してα−APMを製造する方法
の著しく改良された方法に関する。More specifically, α-L-aspartyl-L-phenylalanine is esterified with a medium system consisting of hydrochloric acid and methanol, and the resulting solid α-APM hydrochloride is subjected to solid-liquid separation,
Then, it relates to a significantly improved method of neutralizing the hydrochloride to produce α-APM.
本発明の方法で得られるα−APMは良質の甘味剤を有
し、人工甘味剤として有用な物質であり、近年その需要
は大きいものがある。The α-APM obtained by the method of the present invention has a high-quality sweetener, is a useful substance as an artificial sweetener, and there is a great demand for it in recent years.
（従来の技術） α−APMの製造法に関しては、既に数多くの方法が開示
されているが製造プロセス面からＮ−保護−Ｌ−アスパ
ラギン酸無水物を利用する方法が一般的である。とく
に、Ｌ−アスパラギン酸をギ酸および無水酢酸と反応さ
せることにより１工程で製造可能なＮ−ホルミル−Ｌ−
アスパラギン酸無水物を利用する方法は、原料が容易に
且つ安価に製造でき、その上工程も比較的簡略化される
ことから現状の技術レベルでは工業的製法として適した
方法であると考えられる。(Prior Art) Regarding the production method of α-APM, many methods have already been disclosed, but from the viewpoint of the production process, the method of utilizing N-protected-L-aspartic acid anhydride is general. In particular, N-formyl-L- which can be produced in one step by reacting L-aspartic acid with formic acid and acetic anhydride.
The method using aspartic anhydride is considered to be a method suitable as an industrial production method at the present technical level because the raw material can be easily and inexpensively produced and the process is relatively simplified.
このＮ−ホルミル−Ｌ−アスパラギン酸無水物を用いる
α−APM製造法は、そのほとんどが特開昭46-1350号など
に代表されるように、もう一方の反応原料としてＬ−フ
ェニルアラニンメチルエステルを用いる方法で、Ｎ−ホ
ルミル−α−アスパルチル−Ｌ−フェニルアラニンメチ
ルエステルを中間体として製造した後、保護基のホルミ
ル基を脱離させてα−APMとする技術である。Most of the processes for producing α-APM using this N-formyl-L-aspartic acid anhydride use L-phenylalanine methyl ester as the other reaction raw material, as represented by JP-A-46-1350. It is a technique in which N-formyl-α-aspartyl-L-phenylalanine methyl ester is produced as an intermediate by the method used and then the formyl group of the protecting group is eliminated to obtain α-APM.
そしてＮ−ホルミル−Ｌ−アスパラギン酸無水物とＬ−
フェニルアラニンメチルエステルとの縮合方法、異性体
の抑制方法に関する技術およびホルミル基の脱離方法に
関する技術を中心に色々の方法が提案されている。しか
しながら、このＬ−フェニルアラニンメチルエステルを
一方の反応原料として用いる方法は、Ｌ−フェニルアラ
ニンをエステル化してＬ−フェニルアラニンメチルエス
テルとし、さらにＮ−ホルミル−Ｌ−アスパルギン酸無
水物との縮合反応につなげるまでの工程が繁雑になる。
その上、本発明者らの知見によればＬ−フェニルアラニ
ンメチルエステルが遊離の形態では、溶液中２分子縮合
し、かつ環化して下式構造 のジケトピペラジン化合物に変化し易い性質を有してい
ることがわかった。このことは、工業的には収率の低下
やα−APMの品質劣化等種々のトラブルを誘起する原因
になるものである。And N-formyl-L-aspartic anhydride and L-
Various methods have been proposed, centering on the method for condensation with phenylalanine methyl ester, the technology for suppressing isomers, and the technology for eliminating formyl groups. However, the method of using this L-phenylalanine methyl ester as one of the reaction raw materials is until L-phenylalanine methyl ester is esterified to L-phenylalanine methyl ester, which is further connected to a condensation reaction with N-formyl-L-aspartic acid anhydride. The process becomes complicated.
Furthermore, according to the findings of the present inventors, in the free form of L-phenylalanine methyl ester, two molecules are condensed in a solution and cyclized to give the following structure: It was found that these diketopiperazine compounds have the property of being easily changed. This causes various troubles such as a decrease in yield and deterioration of the quality of α-APM industrially.
したがって、Ｎ−ホルミル−Ｌ−アスパラギン酸無水物
を利用するα−APMの製造法としては、Ｌ−フェニルア
ラニンメチルエステルを使用せずに効率良く製造できる
方法がより好ましい方法と考えられる。Therefore, as a method for producing α-APM using N-formyl-L-aspartic anhydride, a method that can be efficiently produced without using L-phenylalanine methyl ester is considered to be a more preferable method.
Ｌ−フェニルアラニンメチルエステルを使用しないα−
APMの製造法として、Ｎ−ホルミル−Ｌ−アスパラギン
酸無水物を酢酸中、Ｌ−フェニルアラニンと直接縮合さ
せてＮ−ホルミル−α−Ｌ−アスパルチル−Ｌ−フェニ
ルアラニンを製造し、ついでホルミル基を除去してα−
Ｌ−アスパルチル−Ｌ−フェニルアラニンとしたのち、
このα−Ｌ−アスパルチル−Ｌ−フェニルアラニンをメ
タノール中、塩化水素の存在下にエステル化してα−AP
Mを製造する方法（特公昭55-26133号）、及びこのエス
テル化の改良方法としてα−Ｌ−アスパルチル−Ｌ−フ
ェニルアラニンまたはＮ−ホルミル−α−Ｌ−アスパル
チル−Ｌ−フェニルアラニンからその場で生成するα−
Ｌ−アスパルチル−Ｌ−フェニルアラニンを塩化水素、
メタノールおよび水から成る媒体と接触させてエステル
化反応を行い、生成したα−APMを固体状の塩酸塩とし
て析出させて製造する方法（特公昭60-50200号）が開示
されている。しかしながら、前者の方法では２つのカル
ボン酸基のエステル化反応に選択性が低く、目的のα−
APMの他にβ−カルボン酸基がエステル化されたα−Ｌ
−アスパルチル−Ｌ−フェニルアラニン−β−メチルエ
ステルや、ジエステル化されたα−Ｌ−アスパルチル−
Ｌ−フェニルアラニンジメチルエステルが多量に副生
し、その為にα−APMの選択率ならびに収率が低いとい
う欠点がある。これに対して後者の方法は生成したα−
APMが反応の進行に伴い難溶性の塩酸塩として反応系外
に徐々に析出してくるので、比較的好収率でα−APMを
製造しうる方法である。原料の安定性、プロセスの簡便
さ、ならびにα−APMの分離・精製が比較的容易である
点などから考えると、現状のα−APM製造技術のレベル
においては工業的プロセスになりうる技術の一つである
と考えられる。しかしながら、この方法は反応時間が著
しく長いのが難点である。Α-without using L-phenylalanine methyl ester
As a method for producing APM, N-formyl-L-aspartic anhydride is directly condensed with L-phenylalanine in acetic acid to produce N-formyl-α-L-aspartyl-L-phenylalanine, and then the formyl group is removed. Then α-
After making L-aspartyl-L-phenylalanine,
This α-L-aspartyl-L-phenylalanine was esterified in methanol in the presence of hydrogen chloride to obtain α-AP.
A method for producing M (Japanese Examined Patent Publication No. 55-26133), and as an improved method for this esterification, produced in situ from α-L-aspartyl-L-phenylalanine or N-formyl-α-L-aspartyl-L-phenylalanine. Α-
L-aspartyl-L-phenylalanine is hydrogen chloride,
A method (Japanese Patent Publication No. 60-50200) is disclosed in which an esterification reaction is carried out by contacting with a medium consisting of methanol and water, and the produced α-APM is precipitated as a solid hydrochloride salt. However, the former method has low selectivity for the esterification reaction of two carboxylic acid groups, and the desired α-
Α-L in which β-carboxylic acid group is esterified in addition to APM
-Aspartyl-L-phenylalanine-β-methyl ester and diesterified α-L-aspartyl-
A large amount of L-phenylalanine dimethyl ester is produced as a by-product, which has a drawback that the selectivity and yield of α-APM are low. On the other hand, the latter method generates α-
Since APM gradually precipitates out of the reaction system as a hardly soluble hydrochloride with the progress of the reaction, it is a method capable of producing α-APM in a relatively good yield. Considering the stability of raw materials, the simplicity of the process, and the relative ease of separating and purifying α-APM, at the current level of α-APM manufacturing technology, it is one of the technologies that can be an industrial process. It is considered to be one. However, this method has a drawback that the reaction time is extremely long.
この特公昭に記載の実施例の中、Ｎ−ホルミル−α−Ｌ
−アスパルチル−Ｌ−フェニルアラニンからその場で生
成するα−Ｌ−アスパルチル−Ｌ−フェニルアラニンを
用いての例において、反応時間は数日間と長時間を要し
ており、また収率でも反応後単離されたα−APM塩酸塩
を中和して得られる遊離α−APM段階での収率が、およ
そ50〜60％であり、必ずしも満足しうるものではない。Among the examples described in this publication, N-formyl-α-L
-In the example of using α-L-aspartyl-L-phenylalanine which is generated in situ from aspartyl-L-phenylalanine, the reaction time is as long as several days, and the isolation after the reaction is performed even in the yield. The yield in the free α-APM stage obtained by neutralizing the obtained α-APM hydrochloride is about 50 to 60%, which is not always satisfactory.
（発明が解決しようとする問題点） 本発明の課題は、工業的な製造条件で、好収率でα−AP
Mを製造しうる方法を提供することである。(Problems to be Solved by the Invention) An object of the present invention is to produce α-AP in good yield under industrial manufacturing conditions.
It is to provide a method capable of producing M.
（問題点を解決する為の手段） 本発明者らは前記したようなα−APM製造技術の現状を
踏まえ、また溶液中での安定性に問題のあるＬ−フェニ
ルアラニンメチルエステルを用いずに製造できるα−Ｌ
−アスパルチル−Ｌ−フェニルアラニンをメチルエステ
ル化する方法で、さらに効率良くα−APMを製造する方
法を鋭意検討した。とくにα−Ｌ−アスパルチル−Ｌ−
フェニルアラニンを塩酸とメタノールの媒体中にてエス
テル化してさらに高収率に、そして反応時間も短縮しう
る方法を鋭意検討した結果、反応系に塩化マグネシウム
をある濃度で共存させた場合には、無添加の場合に比較
してAPL生成速度を顕著に速めることが可能で、同一の
反応時間では一段と高いα−APM収率が得られることを
見出した。(Means for Solving Problems) Based on the current state of the α-APM production technology as described above, the present inventors have manufactured without using L-phenylalanine methyl ester, which has a problem in stability in a solution. Can α-L
-Aspartyl-L-phenylalanine has been earnestly studied for a method for producing α-APM more efficiently by a method of methyl esterification. Especially α-L-aspartyl-L-
As a result of intensive studies on a method of esterifying phenylalanine in a medium of hydrochloric acid and methanol to further improve the yield and shorten the reaction time, when magnesium chloride coexisted in the reaction system at a certain concentration, no reaction was observed. It was found that the rate of APL formation can be markedly increased as compared with the case of addition, and a higher α-APM yield can be obtained at the same reaction time.
塩化マグネシウム以外の無機塩類についても種々検討し
たが、このような効果は塩化マグネシウム以外の無機塩
ではほとんど認められず、塩化マグネシウムに特有の効
果であることがわかった。Various studies were also carried out on inorganic salts other than magnesium chloride, but it was found that such effects were hardly observed with inorganic salts other than magnesium chloride, and that they were effects specific to magnesium chloride.
図−１は、α−Ｌ−アスパルチル−Ｌ−フェニルアラニ
ンを25％塩酸（3.7モル比）とメタノール（3.75モル
比）とから成る媒体中、20〜25℃でエステル化反応を行
った際に種々の無機塩添加の有無条件下に経時的α−AP
Mの生成率を追跡した結果である。図−１に示されるよ
うに塩酸とメタノールの媒体系のみの場合には反応を３
日間行ってもα−APM生成率は原料のα−Ｌ−アスパル
チル−Ｌ−フェニルアラニンに対しておよそ73モル％で
あるが、塩化マグネシウムをα−Ｌ−アスパルチル−Ｌ
−フェニルアラニンに対して0.41モル％比添加して反応
させた場合には反応２日後で、78モル％となり、３日後
には82モル％まで達する。また塩化マグネシウムの代わ
りに塩化ナトリウムまたは硫酸マグネシウムを添加して
反応を行った場合はAPL生成速度は無添加の場合とほと
んど同じである。Figure 1 shows the results of various esterification reactions of α-L-aspartyl-L-phenylalanine in a medium consisting of 25% hydrochloric acid (3.7 mol ratio) and methanol (3.75 mol ratio) at 20-25 ° C. Α-AP with and without addition of inorganic salts
This is the result of tracking the generation rate of M. As shown in Fig. 1, the reaction is carried out in the case of only the medium system of hydrochloric acid and methanol.
The α-APM production rate was about 73 mol% with respect to the raw material α-L-aspartyl-L-phenylalanine even after the day, but magnesium chloride was mixed with α-L-aspartyl-L.
-When the reaction is carried out by adding 0.41 mol% to phenylalanine, the reaction reaches 78 mol% two days after the reaction and reaches 82 mol% after three days. When the reaction was carried out by adding sodium chloride or magnesium sulfate instead of magnesium chloride, the APL generation rate was almost the same as that without addition.
本発明はこのような知見に基づいて成されたものであ
り、塩化マグネシウムの上記したような効果は従来知ら
れていない。The present invention has been made on the basis of such findings, and the above-described effects of magnesium chloride have not been heretofore known.
本発明は、α−Ｌ−アスパルチル−Ｌ−フェニルアラニ
ンを塩酸およびメタノールから成る媒体中でエステル化
し、生成した固体状のα−APM塩酸塩を固液分離し、こ
の塩酸塩を中和してα−APMを製造する方法において、
エステル化反応を塩化マグネシウムの存在下に行うこと
を特徴とするα−APMの製造法である。In the present invention, α-L-aspartyl-L-phenylalanine is esterified in a medium consisting of hydrochloric acid and methanol, solid α-APM hydrochloride produced is subjected to solid-liquid separation, and the hydrochloride is neutralized to obtain α-APM. -In the method of manufacturing APM,
The method for producing α-APM is characterized in that the esterification reaction is carried out in the presence of magnesium chloride.
そして、更には原料のα−Ｌ−アスパルチル−Ｌ−フェ
ニルアラニンとしては、Ｎ−ホルミル−α−Ｌ−アスパ
ルチル−Ｌ−フェニルアラニンからその場で生成するα
−Ｌ−アスパルチル−Ｌ−フェニルアラニンに本発明の
方法が適用でき、これらの方法も併せて提供するもので
ある。Further, as the raw material α-L-aspartyl-L-phenylalanine, α produced in situ from N-formyl-α-L-aspartyl-L-phenylalanine
The method of the present invention can be applied to -L-aspartyl-L-phenylalanine, and these methods are also provided.
本発明の方法においては、α−Ｌ−アスパルチル−Ｌ−
フェニルアラニンが原料として使用される。このα−Ｌ
−アスパルチル−Ｌ−フェニルアラニンは必ずしも単離
されたものである必要はなく、Ｎ−ホルミル−α−Ｌ−
アスパルチル−Ｌ−フェニルアラニンを塩酸中、または
塩酸とメタノールとの媒体中、高められた温度で処理し
てホルミル基を除去することによってその場で生成され
るものでも良い。In the method of the present invention, α-L-aspartyl-L-
Phenylalanine is used as a raw material. This α-L
-Aspartyl-L-phenylalanine does not necessarily have to be isolated, and N-formyl-α-L-
It may also be produced in situ by treating aspartyl-L-phenylalanine in hydrochloric acid or in a medium of hydrochloric acid and methanol at elevated temperature to remove the formyl group.
原料の一つであるＮ−ホルミル−α−Ｌ−アスパルチル
−Ｌ−フェニルアラニンはＮ−ホルミル−Ｌ−アスパラ
ギン酸無水物とＬ−フェニルアラニンを縮合させること
によって製造できる。とくに水溶媒中で縮合させる特開
昭61-143397号の方法で効率良く製造できる。またα−
Ｌ−アスパルチル−Ｌ−フェニルアラニンはＮ−ホルミ
ル−α−Ｌ−アスパルチル−Ｌ−フェニルアラニンを水
性媒体中、酸と接触させる等の公知のホルミル基除去方
法を利用しても製造できる。これらの原料化合物はＮ−
ホルミル−Ｌ−アスパラギン酸無水物とＬ−フェニルア
ラニンを縮合させる際に副生するβ−異性体、即ち、Ｎ
−ホルミル−β−Ｌ−アスパルチル−Ｌ−フェニルアラ
ニンや原料由来のＮ−ホルミル−Ｌ−アスパラギン酸ま
たはＬ−フェニルアラニンが越ち込されるなら、本発明
の方法に影響しない範囲で混入していても何ら問題な
く、特にβ−異性体は、およそ30重量％程度までは本発
明の方法においてα−APM塩酸塩の析出を極端に妨害し
ないばかりか、このβ−異性体由来の化合物の析出も起
こらず、反応を通して得られるα−APM塩酸塩の品質を
低下させるものではない。One of the raw materials, N-formyl-α-L-aspartyl-L-phenylalanine, can be produced by condensing N-formyl-L-aspartic acid anhydride and L-phenylalanine. Particularly, it can be efficiently produced by the method of JP-A-61-143397 in which condensation is performed in a water solvent. Also α-
L-Aspartyl-L-phenylalanine can also be produced by utilizing a known formyl group removing method such as contacting N-formyl-α-L-aspartyl-L-phenylalanine with an acid in an aqueous medium. These starting compounds are N-
Β-isomer which is a by-product during the condensation of formyl-L-aspartic anhydride and L-phenylalanine, that is, N
-Formyl-β-L-aspartyl-L-phenylalanine and N-formyl-L-aspartic acid or L-phenylalanine derived from raw materials are incorporated, even if they are mixed in a range not affecting the method of the present invention. In particular, the β-isomer does not extremely disturb the precipitation of α-APM hydrochloride in the method of the present invention up to about 30% by weight, and precipitation of a compound derived from this β-isomer also occurs without any problem. Therefore, it does not deteriorate the quality of α-APM hydrochloride obtained through the reaction.
本発明の方法において、α−Ｌ−アスパルチル−Ｌ−フ
ェニルアラニンのエステル化反応は塩酸とメタノールと
から成る媒体中、塩化マグネシウムの存在下に実施され
る。In the method of the present invention, the esterification reaction of α-L-aspartyl-L-phenylalanine is carried out in the presence of magnesium chloride in a medium consisting of hydrochloric acid and methanol.
媒体の塩酸ならびにメタノールはそれぞれ原料のα−Ｌ
−アスパルチル−Ｌ−フェニルアラニンに対して１当量
以上使用され、とくに、塩酸については1.1当量以上用
いるのがα−APM塩酸塩収率の点で好ましい。使用する
塩酸の濃度は５〜33重量％、好適には10〜30重量％の範
囲であり、またメタノールは媒体中のメタノール／H2O
（重量比）で0.03〜0.40、このましくは、0.04〜0.35の
範囲である。塩酸およびメタノールがこれらの範囲外で
あると、α−APM生成速度が著しく緩慢になったり、ま
た生成したα−APMの溶解度が高くなり、その為、α−A
PM塩酸塩が析出し難くなったりして好ましくない。Hydrochloric acid and methanol as media are α-L as raw materials, respectively.
-Aspartyl-L-phenylalanine is used in an amount of 1 equivalent or more, and in particular, hydrochloric acid is preferably used in an amount of 1.1 equivalents or more from the viewpoint of the yield of α-APM hydrochloride. The concentration of hydrochloric acid used is in the range of 5 to 33% by weight, preferably 10 to 30% by weight, and methanol is methanol / H 2 O in the medium.
(Weight ratio) is 0.03 to 0.40, and preferably 0.04 to 0.35. If the amounts of hydrochloric acid and methanol are outside these ranges, the α-APM production rate will be remarkably slow, and the produced α-APM will have a high solubility.
It is not preferable because PM hydrochloride is hard to precipitate.
本発明の方法において、塩酸とメタノールから成る媒体
中に共存させる塩化マグネシウムは、前記媒体中におい
て溶解状態で効果を表すものであり、媒体中の塩化マグ
ネシウム濃度が低すぎるとその効果はほとんどなく、ま
た高すぎると無添加の場合に比較して逆にα−APM生成
速度が遅くなり、場合によってはα−APM塩酸塩が析出
しないこともあり得る。塩化マグネシウムの使用量は、
媒体の塩酸濃度やメタノール濃度によって好適な条件は
変わるが、塩酸とメタノールから成る媒体中での濃度で
0.5〜25重量％、好適には0.7〜20重量％である。共存さ
せる塩化マグネシウムの量が上記の範囲であると、塩化
マグネシウムの共存しない系でのエステル化よりもα−
APM生成速度が速まりその効果が得られる。In the method of the present invention, magnesium chloride coexisting in a medium consisting of hydrochloric acid and methanol represents an effect in a dissolved state in the medium, and there is almost no effect when the concentration of magnesium chloride in the medium is too low, On the other hand, if it is too high, the α-APM generation rate will be slower than in the case of no addition, and in some cases, α-APM hydrochloride may not precipitate. The amount of magnesium chloride used is
Suitable conditions vary depending on the hydrochloric acid concentration and methanol concentration of the medium, but the concentration in the medium consisting of hydrochloric acid and methanol
It is 0.5 to 25% by weight, preferably 0.7 to 20% by weight. When the amount of magnesium chloride to be coexistent is in the above range, α-than the esterification in a system without magnesium chloride coexisting
APM generation speed is increased and its effect is obtained.
本発明の方法でのα−Ｌ−アスパルチル−Ｌ−フェニル
アラニンのエステル化反応は、原料反応媒体および塩化
マグネシウムの装入順序等は特に限定されるものではな
く、例えば、塩酸とメタノールから成る媒体に塩化マグ
ネシウムを装入溶解した溶液中にα−Ｌ−アスパルチル
−Ｌ−フェニルアラニンを装入溶解させ、所定の温度で
反応させることによって達成される。In the esterification reaction of α-L-aspartyl-L-phenylalanine in the method of the present invention, the raw material reaction medium and the charging order of magnesium chloride are not particularly limited, and for example, a medium consisting of hydrochloric acid and methanol is used. This is achieved by charging and dissolving α-L-aspartyl-L-phenylalanine in a solution in which magnesium chloride is charged and dissolved, and reacting at a predetermined temperature.
エステル化反応温度は、通常は０〜50℃、好ましくは10
〜40℃である。低すぎるとエステル化反応速度が遅く、
また高すぎると生成したα−APM塩酸塩の溶解度が高く
なるので、該塩酸塩が析出し難くなってα−APM収率が
低下する。The esterification reaction temperature is usually 0 to 50 ° C, preferably 10
~ 40 ° C. If it is too low, the esterification reaction rate will be slow,
On the other hand, if it is too high, the solubility of the α-APM hydrochloride produced becomes high, so that it becomes difficult for the hydrochloride to precipitate and the α-APM yield decreases.
本発明の方法において、α−Ｌ−アスパルチル−Ｌ−フ
ェニルアラニンは、その前段の化合物であるＮ−ホルミ
ル−α−Ｌ−アスパルチル−Ｌ−フェニルアラニンから
その場で生成するα−Ｌ−アスパルチル−Ｌ−フェニル
アラニンも使用できる。In the method of the present invention, α-L-aspartyl-L-phenylalanine is produced in situ from N-formyl-α-L-aspartyl-L-phenylalanine which is a compound of the preceding stage, α-L-aspartyl-L-. Phenylalanine can also be used.
Ｎ−ホルミル−α−Ｌ−アスパルチル−Ｌ−フェニルア
ラニンからその場でα−Ｌ−アスパルチル−Ｌ−フェニ
ルアラニンを生成させるには、引きつづいて行うエステ
ル化反応で使用される量またはそれ以上の量の塩酸中で
Ｎ−ホルミル−α−Ｌ−アスパルチル−Ｌ−フェニルア
ラニンを加熱することによって達成される。この際、エ
ステル化工程で用いる量またはそれより少ない量のメタ
ノールを共存させて行うことも可能である。加熱される
温度はペプチドの開裂等の副反応を抑制する上で65℃以
下が良く、好ましくは40〜60℃である。Ｎ−ホルミル−
α−Ｌ−アスパルチル−Ｌ−フェニルアラニンからα−
Ｌ−アスパルチル−Ｌ−フェニルアラニンの生成をメタ
ノールの共存下に行った場合には、塩酸濃度ならびにメ
タノール濃度によってその程度は変化するが生成したα
−Ｌ−アスパルチル−Ｌ−フェニルアラニンはそのエス
テル化合物との混合物として生成するのは勿論のことで
ある。このようにして生成したα−Ｌ−アスパルチル−
Ｌ−フェニルアラニンは媒体の塩酸量や濃度ならびにメ
タノール量などを調整したのち、所定量の塩化マグネシ
ウムを装入溶解して、エステル化することによりα−AP
M塩酸塩が製造される。In-situ production of α-L-aspartyl-L-phenylalanine from N-formyl-α-L-aspartyl-L-phenylalanine can be achieved by using an amount of the amount used in the subsequent esterification reaction or higher. This is accomplished by heating N-formyl-α-L-aspartyl-L-phenylalanine in hydrochloric acid. At this time, it is also possible to coexist with the amount of methanol used in the esterification step or a smaller amount thereof. The heating temperature is preferably 65 ° C. or lower, preferably 40 to 60 ° C., in order to suppress side reactions such as peptide cleavage. N-formyl-
α-L-aspartyl-L-phenylalanine to α-
When L-aspartyl-L-phenylalanine was produced in the coexistence of methanol, α was produced depending on the concentration of hydrochloric acid and the concentration of methanol.
Of course, -L-aspartyl-L-phenylalanine is produced as a mixture with its ester compound. Α-L-aspartyl-produced in this way
L-phenylalanine is prepared by adjusting the amount and concentration of hydrochloric acid in the medium and the amount of methanol, then charging and dissolving a prescribed amount of magnesium chloride and esterifying it to form α-AP.
M hydrochloride is produced.
本発明においては生成したα−APMの塩酸塩は逐次反応
系より結晶として析出する。従ってエステル化反応後は
必要に応じて冷却後、円心分離等の濾過操作によってα
−APM塩酸塩を取得できる。この塩酸塩は常法により、
例えば、水中で水酸化アルカリ、炭酸アルカリまたは重
炭酸アルカリなどの塩基で中和することによって遊離の
α−APMに変換できる。In the present invention, the produced hydrochloride of α-APM is precipitated as crystals from the sequential reaction system. Therefore, after the esterification reaction, if necessary, after cooling, the α
-APM hydrochloride can be obtained. This hydrochloride is
For example, it can be converted into free α-APM by neutralizing in water with a base such as an alkali hydroxide, an alkali carbonate or an alkali bicarbonate.
（効果） 本発明の方法によれば、α−Ｌ−アスパルチル−Ｌ−フ
ェニルアラニンを塩酸とメタノールとから成る媒体中で
エステル化反応を行いα−APM塩酸塩を製造するに際し
て、工業的にも安価な塩化マグネシウムの添加のみでα
−APM塩酸塩の生成速度が顕著に速まり、より高収率で
α−APMが製造できる。またα−Ｌ−アスパルチル−Ｌ
−フェニルアラニンは必ずしも単離されたものである必
要はなく、その前段の化合物であるＮ−ホルミル−α−
Ｌ−アスパルチル−Ｌ−フェニルアラニンからその場で
生成されるものであってもよく、エステル化法によるα
−APMの優れた改良法と言える。(Effect) According to the method of the present invention, α-L-aspartyl-L-phenylalanine undergoes an esterification reaction in a medium consisting of hydrochloric acid and methanol to produce α-APM hydrochloride, which is industrially inexpensive. Α by adding pure magnesium chloride
-The production rate of APM hydrochloride is remarkably increased, and α-APM can be produced in a higher yield. In addition, α-L-aspartyl-L
-Phenylalanine does not necessarily have to be isolated, and the compound of the preceding stage, N-formyl-α-
It may be generated in-situ from L-aspartyl-L-phenylalanine, and may be produced by the esterification method.
-It can be said that it is an excellent improvement method of APM.
（実施例） 以下、実施例により本発明を詳細に説明する。尚、実施
例中の高速液体のクロマトグラフィーの分析条件は以下
の通りである。(Examples) Hereinafter, the present invention will be described in detail with reference to Examples. The analytical conditions for the chromatography of the high performance liquid in the examples are as follows.
高速液体クロマトグラフィーの分析条件 カラム YMC pack A−312 6mmφ×150mm （充填剤 ODS） 移動相 0.005M/l ヘプタンスルホン酸ナトリウム水溶
液：メタノール＝65:35（体積比） （リン酸でpH＝2.5に調整） 流 量 1ml/min 検出器 紫外分光光度計 実施例１ 35％塩酸38.6g、水11gおよびメタノール12.0gから調整
された媒体中に塩化マグネシウム６水和物8.3gを溶解し
た（塩化マグネシウムの媒体中での濃度は5.6重量
％）。この溶液中にα−Ｌ−アスパルチル−Ｌ−フェニ
ルアラニン28.0gを室温で溶解した。その後20〜25℃で
３日間（72Hr）反応させた。α−APMの生成率を高速液
体クロマトグラフィーで追跡した結果は、図−１に示
す。この結果から３日後のα−APM生成率は81.9％（対
α−Ｌ−アスパルチル−Ｌ−フェニルアラニン）であっ
た。Analytical conditions for high performance liquid chromatography Column YMC pack A-312 6mmφ x 150mm (packing ODS) Mobile phase 0.005M / l sodium heptane sulfonate aqueous solution: methanol = 65:35 (volume ratio) (pH = 2.5 with phosphoric acid) Adjustment) Flow rate 1 ml / min Detector UV spectrophotometer Example 1 8.3 g of magnesium chloride hexahydrate was dissolved in a medium prepared from 35% hydrochloric acid 38.6 g, water 11 g and methanol 12.0 g (of magnesium chloride). The concentration in the medium is 5.6% by weight). 28.0 g of α-L-aspartyl-L-phenylalanine was dissolved in this solution at room temperature. Then, the mixture was reacted at 20 to 25 ° C for 3 days (72 hours). Figure 1 shows the results of high-performance liquid chromatography tracing of α-APM production rate. From this result, the α-APM production rate after 3 days was 81.9% (vs α-L-aspartyl-L-phenylalanine).
その後５℃に冷却し、析出している結晶を吸引濾過し、
５℃以下に冷却された１規定塩酸で洗浄することにより
α−APM塩酸塩の湿ケーキを得た。高速液体クロマトグ
ラフィーで分析の結果、23.3gのα−APM（遊離換算）を
含有していた。収率79.2％（対α−Ｌ−アスパルチル−
Ｌ−フェニルアラニン） ここに得たα−APM塩酸塩の湿ケーキを水400ml中、常法
により20％炭酸ナトリウム水溶液でpH5.2に中和し、５
℃に冷却後濾過、水洗ののち減圧下に乾燥することによ
り20.6gのα−APMを得た。After that, it was cooled to 5 ° C., and the precipitated crystals were suction filtered,
A wet cake of α-APM hydrochloride was obtained by washing with 1N hydrochloric acid cooled to 5 ° C or lower. As a result of analysis by high performance liquid chromatography, it contained 23.3 g of α-APM (free conversion). Yield 79.2% (vs. α-L-aspartyl-
L-phenylalanine) The wet cake of α-APM hydrochloride obtained here was neutralized to a pH of 5.2 with 20% sodium carbonate aqueous solution in 400 ml of water by a conventional method,
The mixture was cooled to ℃, filtered, washed with water, and dried under reduced pressure to obtain 20.6 g of α-APM.
収率70.0％（対α−Ｌ−アスパルチル−Ｌ−フェニルア
ラニン） 〔α〕20＝＋15.7°（Ｃ＝4 15規定ギ酸） 実施例２ 実施例１において水を14.5gにまた塩化マグネシウム６
水和物を1.6gに代える以外は実施例１と同様に反応を行
った。α−APMの生成率を高速液体クロマトグラフィー
で追跡した結果は、図−１に示す。３日後のα−APM生
成率は79.4％（対α−Ｌ−アスパルチル−Ｌ−フェニル
アラニン）であり、また実施例１と同じようにして単離
されたα−APM塩酸塩の単離収率は、76.9％（対α−Ｌ
−アスパルチル−Ｌ−フェニルアラニン）であった。Yield 70.0% (relative to α-L-aspartyl-L-phenylalanine) [α] 20 = + 15.7 ° (C = 415 normal formic acid) Example 2 In Example 1, water was added to 14.5 g and magnesium chloride 6
The reaction was performed in the same manner as in Example 1 except that the hydrate was replaced with 1.6 g. Figure 1 shows the results of high-performance liquid chromatography tracing of α-APM production rate. The production rate of α-APM after 3 days was 79.4% (vs. α-L-aspartyl-L-phenylalanine), and the isolated yield of α-APM hydrochloride isolated in the same manner as in Example 1 was. , 76.9% (vs α-L
-Aspartyl-L-phenylalanine).
比較例１ 実施例１において塩化マグネシウムを添加しない他は実
施例１と同様にエステル化反応を行った（但し水の量を
15.4gとし塩酸濃度を実施例１と同じく25％濃度にし
た）。α−APMの生成率を高速液体クロマトグラフィー
で追跡した結果は、図−１に示す。20〜25℃、３日間反
応後のα−APM生成率は73.0％（対α−Ｌ−アスパルチ
ル−Ｌ−フェニルアラニン）であった。また実施例１と
同じようにして単離されたα−APM塩酸塩の単離収率は6
9.8％（対α−Ｌ−アスパルチル−Ｌ−フェニルアラニ
ン）であった。Comparative Example 1 The esterification reaction was carried out in the same manner as in Example 1 except that magnesium chloride was not added.
The concentration of hydrochloric acid was set to 15.4 g and the concentration of hydrochloric acid was set to 25% as in Example 1). Figure 1 shows the results of high-performance liquid chromatography tracing of α-APM production rate. The α-APM production rate after reaction at 20 to 25 ° C for 3 days was 73.0% (vs α-L-aspartyl-L-phenylalanine). The isolated yield of α-APM hydrochloride isolated in the same manner as in Example 1 was 6
9.8% (vs. α-L-aspartyl-L-phenylalanine).
比較例２〜３ 実施例１において塩化マグネシウムの代わりに塩化ナシ
リウム（比較例２）4.8g、或いは無水硫酸マグネシウム
（比較例３）4.9gを用いる以外は水の量を実施例１に合
わせて同様に反応を行った。反応３日後のα−APM生成
率はそれぞれ73.6％、72.1％（対α−Ｌ−アスパルチル
−Ｌ−フェニルアラニン）で比較例１とほとんど差はな
かった。Comparative Examples 2 to 3 Same as Example 1 except that 4.8 g of Nasium chloride (Comparative Example 2) or 4.9 g of anhydrous magnesium sulfate (Comparative Example 3) was used instead of magnesium chloride in Example 1. Was reacted. The α-APM production rates after 3 days of reaction were 73.6% and 72.1% (vs. α-L-aspartyl-L-phenylalanine), which were almost the same as those of Comparative Example 1.
比較例４ 実施例２において塩化マグネシウム６水和物の使用量を
0.6gに代える他は実施例２と同様にエステル化反応を行
った結果、同じく反応３日後のα−APM生成率は73.9％
（対α−Ｌ−アスパルチル−Ｌ−フェニルアラニン）で
あった。Comparative Example 4 The amount of magnesium chloride hexahydrate used in Example 2 was changed to
As a result of carrying out the esterification reaction in the same manner as in Example 2 except that the amount was changed to 0.6 g, the rate of α-APM production after 3 days was 73.9%.
(Vs. α-L-aspartyl-L-phenylalanine).
実施例３ 35％HCl 41.7g、水39.4gおよびメタノール9.6gから成る
媒体中に無水塩化マグネシウム10.0gを溶解し、さらに
α−Ｌ−アスパルチル−Ｌ−フェニルアラニン28.0gを
溶解した。その後30〜35℃で３日間反応させた。高速液
体クロマトグラフィーにてα−APM生成率を分析の結果8
1.2％（対α−Ｌ−アスパルチル−Ｌ−フェニルアラニ
ン）であった。Example 3 10.0 g of anhydrous magnesium chloride was dissolved in a medium composed of 41.7 g of 35% HCl, 39.4 g of water and 9.6 g of methanol, and 28.0 g of α-L-aspartyl-L-phenylalanine was further dissolved. Then, the mixture was reacted at 30 to 35 ° C for 3 days. Results of analysis of α-APM production rate by high performance liquid chromatography 8
It was 1.2% (vs. α-L-aspartyl-L-phenylalanine).
比較例５ 実施例３において塩化マグネシウムを添加せずに反応を
行った。３日後のα−APM生成率は74.6％であった。Comparative Example 5 The reaction was carried out in Example 3 without adding magnesium chloride. The α-APM production rate after 3 days was 74.6%.
実施例４ ４規定塩酸60.8gとメタノール12gとから成る媒体を50℃
に昇温し50〜55℃でＮ−ホルミル−α−Ｌ−アスパルチ
ル−Ｌ−フェニルアラニン30.8gをおよそ30分要して加
えた。その後50〜60℃で２時間反応させてホルミル基を
除去した。室温まで冷却後35％HCl 17.7gと無水塩化マ
グネシウム7.8gを装入した。20〜25℃で４日間反応させ
たところα−APM生成率は82.5％（対α−Ｌ−アスパル
チル−Ｌ−フェニルアラニン）であった。５℃に冷却後
吸引濾過し、５℃以下に冷却された１規定塩酸で洗浄す
ることにより23.4gのα−APM（遊離換算）を含有するα
−APM塩酸塩のケーキを得た。収率:79.5％（対α−Ｌ−
アスパルチル−Ｌ−フェニルアラニン） 比較例６ 実施例４において塩化マグネシウムを添加せずに同様に
反応を行った。20〜25℃４日間反応後α−APM生成率は7
4.3％（対α−Ｌ−アスパルチル−Ｌ−フェニルアラニ
ン）であった。Example 4 A medium composed of 60.8 g of 4N hydrochloric acid and 12 g of methanol was treated at 50 ° C.
The temperature was raised to 50 ° C. to 55 ° C., and 30.8 g of N-formyl-α-L-aspartyl-L-phenylalanine was added over about 30 minutes. After that, the formyl group was removed by reacting at 50 to 60 ° C. for 2 hours. After cooling to room temperature, 17.7 g of 35% HCl and 7.8 g of anhydrous magnesium chloride were charged. When reacted at 20 to 25 ° C for 4 days, the α-APM production rate was 82.5% (vs α-L-aspartyl-L-phenylalanine). After cooling to 5 ° C, suction filtration, and washing with 1N hydrochloric acid cooled to 5 ° C or below, α containing 23.4g of α-APM (free conversion)
-A cake of APM hydrochloride was obtained. Yield: 79.5% (vs α-L-
Aspartyl-L-phenylalanine) Comparative Example 6 The same reaction as in Example 4 was performed without adding magnesium chloride. After reacting at 20-25 ℃ for 4 days, α-APM production rate is 7
It was 4.3% (vs. α-L-aspartyl-L-phenylalanine).
図−１は、実施例１、実施例２および比較例１のα−AP
M生成率と時間の関係を示すものである。 図中の符号はそれぞれつぎの通りである。 実施例１（MgCl20.41モル比添加） 実施例２（MgCl20.08モル比添加） 比較例１（無機塩無添加）FIG. 1 shows α-AP of Examples 1, 2 and Comparative Example 1.
It shows the relationship between the M generation rate and time. The symbols in the figure are as follows. Example 1 (MgCl 2 0.41 molar ratio added) Example 2 (MgCl 2 0.08 molar ratio added) Comparative Example 1 (no inorganic salt added)
Claims (3)
ニンを塩酸およびメタノールから成る媒体中でエステル
化し、生成した固体状のα−Ｌ−アスパルチル−Ｌ−フ
ェニルアラニンメチルエステルの塩酸塩を固液分離し、
該塩酸塩を中和することから成るα−Ｌ−アスパルチル
−Ｌ−フェニルアラニンメチルエステルの製造法におい
て、エステル化反応を塩化マグネシウムの存在下に行う
ことを特徴とするα−Ｌ−アスパルチル−Ｌ−フェニル
アラニンメチルエステルの製造法。1. α-L-Aspartyl-L-phenylalanine is esterified in a medium consisting of hydrochloric acid and methanol, and the resulting solid salt of α-L-aspartyl-L-phenylalanine methyl ester is subjected to solid-liquid separation. ,
In a method for producing α-L-aspartyl-L-phenylalanine methyl ester, which comprises neutralizing the hydrochloride, α-L-aspartyl-L-, characterized in that the esterification reaction is carried out in the presence of magnesium chloride. Process for producing phenylalanine methyl ester.
ニンが、Ｎ−ホルミル−α−Ｌ−アスパルチル−Ｌ−フ
ェニルアラニンからその場で生成するα−Ｌ−アスパル
チル−Ｌ−フェニルアラニンである特許請求の範囲第１
項記載の方法。2. An α-L-aspartyl-L-phenylalanine is an α-L-aspartyl-L-phenylalanine formed in situ from N-formyl-α-L-aspartyl-L-phenylalanine. First
Method described in section.
ルから成る媒体中におよそ0.5〜25重量％の範囲で存在
する特許請求の範囲第１項ならびに第２項記載の方法。3. A process according to claims 1 and 2 wherein magnesium chloride is present in the medium consisting of hydrochloric acid and methanol in the range of approximately 0.5 to 25% by weight.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61305329A JPH07116228B2 (en) | 1986-12-23 | 1986-12-23 | Process for producing α-L-aspartyl-L-phenylalanine methyl ester |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61305329A JPH07116228B2 (en) | 1986-12-23 | 1986-12-23 | Process for producing α-L-aspartyl-L-phenylalanine methyl ester |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63159363A JPS63159363A (en) | 1988-07-02 |
| JPH07116228B2 true JPH07116228B2 (en) | 1995-12-13 |
Family
ID=17943802
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61305329A Expired - Lifetime JPH07116228B2 (en) | 1986-12-23 | 1986-12-23 | Process for producing α-L-aspartyl-L-phenylalanine methyl ester |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH07116228B2 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7635490B2 (en) | 2001-09-28 | 2009-12-22 | Mcneil-Ppc, Inc. | Modified release dosage form |
-
1986
- 1986-12-23 JP JP61305329A patent/JPH07116228B2/en not_active Expired - Lifetime
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7635490B2 (en) | 2001-09-28 | 2009-12-22 | Mcneil-Ppc, Inc. | Modified release dosage form |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS63159363A (en) | 1988-07-02 |
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