JPS6218559B2 - - Google Patents
Info
- Publication number
- JPS6218559B2 JPS6218559B2 JP53111780A JP11178078A JPS6218559B2 JP S6218559 B2 JPS6218559 B2 JP S6218559B2 JP 53111780 A JP53111780 A JP 53111780A JP 11178078 A JP11178078 A JP 11178078A JP S6218559 B2 JPS6218559 B2 JP S6218559B2
- Authority
- JP
- Japan
- Prior art keywords
- ornithine
- hydroxyarginine
- lysine
- threonine
- polypeptide
- 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
Links
- WBVFBXZGHGWBRE-RZOMMOEVSA-N (2r)-2-[[(2s)-6-amino-2-[[(2r)-5-amino-2-[[(2s,3r)-2-[[(2r)-5-amino-2-[[(2s,4s)-2-[[(2s)-2-amino-5-(diaminomethylideneamino)pentanoyl]amino]-5-(diaminomethylideneamino)-4-hydroxypentanoyl]amino]pentanoyl]amino]-3-hydroxybutanoyl]amino]pentanoyl]amino]hexa Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](C[C@H](O)CN=C(N)N)C(=O)N[C@H](CCCN)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](CCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(O)=O)CC1=CC=C(O)C=C1 WBVFBXZGHGWBRE-RZOMMOEVSA-N 0.000 claims description 32
- DETXZQGDWUJKMO-UHFFFAOYSA-N 2-hydroxymethanesulfonic acid Chemical compound OCS(O)(=O)=O DETXZQGDWUJKMO-UHFFFAOYSA-N 0.000 claims description 23
- 150000001413 amino acids Chemical group 0.000 claims description 11
- 239000004472 Lysine Substances 0.000 claims description 10
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 claims description 7
- 239000003910 polypeptide antibiotic agent Substances 0.000 claims description 7
- AHLPHDHHMVZTML-BYPYZUCNSA-N L-Ornithine Chemical compound NCCC[C@H](N)C(O)=O AHLPHDHHMVZTML-BYPYZUCNSA-N 0.000 claims description 6
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 6
- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 claims description 6
- UTJLXEIPEHZYQJ-UHFFFAOYSA-N Ornithine Natural products OC(=O)C(C)CCCN UTJLXEIPEHZYQJ-UHFFFAOYSA-N 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 229960003104 ornithine Drugs 0.000 claims description 6
- 229920001184 polypeptide Polymers 0.000 claims description 6
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 6
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 6
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 6
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 5
- 229910052783 alkali metal Inorganic materials 0.000 claims description 5
- 239000000470 constituent Substances 0.000 claims description 5
- WQFIAVZQVYASHZ-BYPYZUCNSA-N (2s)-5-(diaminomethylideneamino)-2-(hydroxyamino)pentanoic acid Chemical compound NC(=N)NCCC[C@H](NO)C(O)=O WQFIAVZQVYASHZ-BYPYZUCNSA-N 0.000 claims description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 claims description 4
- 150000001340 alkali metals Chemical class 0.000 claims description 4
- 125000003277 amino group Chemical group 0.000 claims description 4
- -1 hydroxyarginine-ornithine-threonine-ornithine-lysine Chemical compound 0.000 claims description 3
- 239000002262 Schiff base Substances 0.000 claims description 2
- 150000004753 Schiff bases Chemical class 0.000 claims description 2
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 claims description 2
- 238000006243 chemical reaction Methods 0.000 description 13
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 238000000034 method Methods 0.000 description 7
- KKVTYAVXTDIPAP-UHFFFAOYSA-M sodium;methanesulfonate Chemical compound [Na+].CS([O-])(=O)=O KKVTYAVXTDIPAP-UHFFFAOYSA-M 0.000 description 7
- 238000003756 stirring Methods 0.000 description 6
- 150000001875 compounds Chemical class 0.000 description 5
- 150000003839 salts Chemical class 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 239000004475 Arginine Substances 0.000 description 4
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 3
- 239000004473 Threonine Substances 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 229910052500 inorganic mineral Inorganic materials 0.000 description 3
- 238000010253 intravenous injection Methods 0.000 description 3
- 239000011707 mineral Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 241000223250 Metarhizium anisopliae Species 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000001093 anti-cancer Effects 0.000 description 2
- 239000012458 free base Substances 0.000 description 2
- 230000000704 physical effect Effects 0.000 description 2
- 230000001766 physiological effect Effects 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 2
- OQCZIHUBJQDIQI-ZCFIWIBFSA-N (2r)-2-amino-5-(diaminomethylideneamino)-2-hydroxypentanoic acid Chemical compound NC(=N)NCCC[C@@](N)(O)C(O)=O OQCZIHUBJQDIQI-ZCFIWIBFSA-N 0.000 description 1
- WFIYPADYPQQLNN-UHFFFAOYSA-N 2-[2-(4-bromopyrazol-1-yl)ethyl]isoindole-1,3-dione Chemical compound C1=C(Br)C=NN1CCN1C(=O)C2=CC=CC=C2C1=O WFIYPADYPQQLNN-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 241000223201 Metarhizium Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000840267 Moma Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 208000000474 Poliomyelitis Diseases 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000555745 Sciuridae Species 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000005907 cancer growth Effects 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 159000000001 potassium salts Chemical class 0.000 description 1
- SSOLNOMRVKKSON-UHFFFAOYSA-N proguanil Chemical compound CC(C)\N=C(/N)N=C(N)NC1=CC=C(Cl)C=C1 SSOLNOMRVKKSON-UHFFFAOYSA-N 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- QYSXJUFSXHHAJI-YRZJJWOYSA-N vitamin D3 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-YRZJJWOYSA-N 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Compounds Of Unknown Constitution (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Description
本発明は、ペプチド性抗生物質である、アルギ
ニン−γ−ヒドロキシアルギニン−オルニチン−
スレオニン−オルニチン−リジンで示されるアミ
ノ酸配列を有するポリペプチド（以下、K582M
−Ａと称する。）のホルムアルデヒド重亜硫酸塩
付加体の製造方法に関する。
既に本発明者は、メタリジウム属に属する菌
株、例えばメタリジウム・アニソプリエ・（メツ
シユ）・ソロク・バール・アニソプリエ582M
（Metarhizium anisopliae（Metsch）Sorok.rar.
anisopliae 582M）株（微工研菌寄第4217号）を
培養して得られる培養液から、各種真菌の発育阻
止、特にカンジダおよび酵母類に有効で、かつ、
ポリオ、インフルエンザ等のウイルス感染の抑
制、更に実験動物がんの発育阻止に有効な物質が
含まれていること及びこの有効物質がK582M−
Ａ及びK582M−Ｂであることを見い出し、この
知見に基づきK582M−Ａ，K582M−Ｂ及びこれ
らの製造法につき特許出願した（特願昭52−
157416号）。
ここにK582M−Ａとは、アルギニン１、スレ
オニン１、チロシン１、オルニチン２、ヒドロキ
シアルギニン１、リジン１を構成アミノ酸とする
直鎖状のペプチド性抗生物質又はその塩をいい、
又、K582M−Ｂとは、アルギニン１、スレオニ
ン１、チロシン１、オルニチン１、ヒドロキシア
ルギニン２を構成アミノ酸とする直鎖状のペプチ
ド性抗生物質又はその鉱酸塩をいう。
本発明者は、上記のペプチド性抗生物質の研究
を発展させるべくその誘導体の開発に鋭意検討を
加えた結果、新規化合物であるK582M−Ａのホ
ルムアルデヒド重亜硫酸塩付加体の合成に成功
し、又、合成された該新規化合物がK582M−Ａ
の有する抗菌活性を減損することなくK582M−
Ａの毒性を低減できることを見い出し本発明を完
成するに至つた。
本発明は、一般式
Ｘ（−NH−CH2−SO3M）4
（式中、Ｘは、アルギニン１、スレオニン１、
チロシン１、オルニチン２、ヒドロキシアルギニ
ン１、リジン１を構成アミノ酸とする直鎖状ペプ
チド性抗生物質であるK582M−Ａから４個の遊
離アミノ基を除いた母核を示し、Ｍはアルカリ金
属又はアンモニウムイオンを示す。）
で示されるK582M−Ａのホルムアルデヒド重亜
硫酸塩付加体に関する。
又、本発明は、一般式
Ｘ（−NH2）4
（式中、Ｘは、アルギニン１、スレオニン１、
チロシン１、オルニチン２、ヒドロキシアルギニ
ン１、リジン１を構成するアミノ酸とする直鎖状
ペプチド性抗生物質であるK582M−Ａから４個
の遊離アミノ基を除いた母核を示す。）
で示されるK582M−Ａに４倍モル量のホルムア
ルデヒドを反応させ、得られたシツフ塩基に４倍
モル量の重亜硫酸塩と反応させるか又はK582M
−Ａに予めホルムアルデヒドと重亜硫酸塩との反
応により得られた、４倍モル量のホルムアルデヒ
ドの重亜硫酸塩付加体を反応させることを特徴と
する、一般式
Ｘ（−NH−CH2−SO3M）4
（式中、Ｘは上で定義した通りであり、Ｍはア
ルカリ金属又はアンモニウムイオンを示す。）
で示されるK582M−Ａのホルムアルデヒド重亜
硫酸塩付加体の製造方法に関するものでもある。
以下に反応式を挙げて本発明のK582M−Ａの
ホルムアルデヒド重亜硫酸塩付加体の製造方法を
説明する。
方法 １
Ｘ（−NH2）4＋4HCHO →Ｘ（−NH＝CH2）4
Ｘ（−Ｎ＝CH2）4＋4MHSO3
→Ｘ（−NH−CH2−SO3M）4
方法 ２
HCHO＋MHSO3→MSO3CH2OH
Ｘ（−NH2）4＋4MSO3CH2OH
→Ｘ（−NH−CH2−SO3M）4
（式中、ＸおよびＭは、上で定義した通りであ
る。）
本発明において原料であるK582M−Ａは、遊
離塩基の形で又はその鉱酸塩（例えば塩酸塩、硫
酸塩など）の形で反応容器に仕込まれる。更に
又、使用される重亜硫酸塩としては、重亜硫酸の
アルカリ金属塩（ナトリウム塩、カリウム塩
等）、アンモニウム塩等の可溶性塩が挙げられ
る。
方法１を実施するには、例えばK582M−Ａ
（遊離塩基又は鉱酸塩）を５〜20％の濃度になる
ように水に溶解し、これに撹拌下ホルムアルデヒ
ドを加える。これにアルカリを加えてPHを7.5〜
9.0に調整し、K582M−Ａのアルデヒド付加体を
完全に沈殿させる。次いでこの付加体を少量の水
に懸濁させ、30〜50℃で撹拌しながら重亜硫酸塩
を加えて反応させれば、K582M−Ａのホルムア
ルデヒド重亜硫酸塩付加体が得られる。
また方法２を実施するには、あらかじめホルム
アルデヒド化合物と重亜硫酸塩とを30〜40℃で撹
拌してホルムアルデヒドの重亜硫酸塩を製造し、
これにK582M−Ａを徐々に加えて反応させる。
斯くして得られたK582M−Ａのホルムアルデ
ヒド重亜硫酸塩付加体を含む反応液は、例えばセ
フアデツクスカラムを用いてゲル過を行ない、
その活性区分を採取して凍結乾燥すれば精製され
たK582M−Ａのホルムアルデヒド重亜硫酸塩付
加体が得られる。
上記の方法１、方法２に基づき実施例１，２の
如くして製造された本発明の化合物の一例である
K582M−Ａのメタンスルホン酸ナトリウムの物
性及び生理的性質は、以下の如くである。
(a) 物性
(1) 性 状 白色無晶形酸性物質
(2) 分解点 202〜210℃で徐々に分解
(3) 比旋光度 〔Ｘ〕３０ Ｄ＝＋1.2（１％H2O）
(4) 元素分析値 C37.4％
H5.6％
N15.5％
(5) 分子式 C45H78N16O23S4Na4
(6) 分子量 1400（蒸気圧法）
(7) 呈色反応 ミロン反応、坂口反応、キサン
トプロテイン反応陽性、ニンヒドリン（中
性）反応、モーリツシユ、トーレンス反応陰
性
(8) 構成アミノ酸
（6N塩酸で封管中110℃で24時間加水分解
して測定した。）
スレオニン１、チロシン１、オルニチン２、
リジン１、アルギニン１、ヒドロキシルアル
ギニン１
(9) 赤外線吸収スペクトル 第１図
(b) 生理的性質
(1) 抗菌活性（最小阻止濃度mcg／ml）
The present invention provides a peptide antibiotic, arginine-γ-hydroxyarginine-ornithine-
A polypeptide having an amino acid sequence of threonine-ornithine-lysine (hereinafter referred to as K582M
-Referred to as A. ) relates to a method for producing a formaldehyde bisulfite adduct. The present inventor has already discovered strains belonging to the genus Metarhizium, such as Metarhizium anisopliae (Metsushiyu) Sorok var anisopliae 582M.
(Metarhizium anisopliae (Metsch) Sorok.rar.
anisopliae 582M) strain (Feikoken Bacteria No. 4217), which is effective in inhibiting the growth of various fungi, especially Candida and yeast, and
Contains a substance that is effective in suppressing viral infections such as polio and influenza, as well as inhibiting the growth of cancer in laboratory animals, and that this effective substance is K582M-
Based on this knowledge, we filed a patent application for K582M-A, K582M-B and their manufacturing method (patent application filed in 1972).
No. 157416). Here, K582M-A refers to a linear peptide antibiotic or a salt thereof whose constituent amino acids are 1 arginine, 1 threonine, 1 tyrosine, 2 ornithine, 1 hydroxyarginine, and 1 lysine.
Further, K582M-B refers to a linear peptide antibiotic or its mineral acid salt having 1 arginine, 1 threonine, 1 tyrosine, 1 ornithine, and 2 hydroxyarginine as constituent amino acids. As a result of intensive studies into the development of derivatives of the above-mentioned peptide antibiotics, the present inventor succeeded in synthesizing a formaldehyde bisulfite adduct of K582M-A, which is a new compound. , the synthesized new compound is K582M-A
K582M− without impairing its antibacterial activity.
They discovered that the toxicity of A can be reduced and completed the present invention. The present invention has the general formula X(-NH- CH2 - SO3M ) 4 (wherein,
This represents the core obtained by removing four free amino groups from K582M-A, a linear peptide antibiotic whose constituent amino acids are 1 tyrosine, 2 ornithine, 1 hydroxyarginine, and 1 lysine, and M is an alkali metal or ammonium. Indicates an ion. ) Regarding the formaldehyde bisulfite adduct of K582M-A shown in The present invention also provides a compound having the general formula X(-NH 2 ) 4 (wherein,
This figure shows the core of K582M-A, a linear peptide antibiotic whose amino acids are 1 tyrosine, 2 ornithine, 1 hydroxyarginine, and 1 lysine, with four free amino groups removed. ) is reacted with 4 times the molar amount of formaldehyde, and the resulting Schiff base is reacted with 4 times the molar amount of bisulfite, or K582M-A is reacted with 4 times the molar amount of bisulfite.
General formula X (-NH-CH 2 -SO 3 The present invention also relates to a method for producing a formaldehyde bisulfite adduct of K582M-A represented by M) 4 (wherein X is as defined above and M represents an alkali metal or ammonium ion). The method for producing the formaldehyde bisulfite adduct of K582M-A of the present invention will be described below with reference to the reaction formula. Method 1 X(-NH 2 ) 4 +4HCHO →X(-NH=CH 2 ) 4 X(-N=CH 2 ) 4 +4MHSO 3
→X(-NH-CH 2 -SO 3 M) 4 Method 2 HCHO+MHSO 3 →MSO 3 CH 2 OH X(-NH 2 ) 4 +4MSO 3 CH 2 OH
→X(-NH- CH2 - SO3M ) 4 (wherein, X and M are as defined above) It is charged to the reaction vessel in the form of mineral acid salts (e.g. hydrochloride, sulfate, etc.). Furthermore, examples of the bisulfite used include soluble salts of bisulfite such as alkali metal salts (sodium salts, potassium salts, etc.) and ammonium salts. To implement method 1, for example, K582M-A
(free base or mineral acid salt) in water to a concentration of 5-20%, and formaldehyde is added to this with stirring. Add alkali to this and adjust the pH to 7.5~
9.0 to completely precipitate the aldehyde adduct of K582M-A. Next, this adduct is suspended in a small amount of water, and bisulfite is added and reacted with stirring at 30 to 50°C to obtain a formaldehyde bisulfite adduct of K582M-A. In addition, in order to carry out method 2, the formaldehyde compound and bisulfite are stirred in advance at 30 to 40°C to produce formaldehyde bisulfite,
K582M-A is gradually added to this and reacted. The reaction solution containing the formaldehyde bisulfite adduct of K582M-A thus obtained is subjected to gel filtration using, for example, a Sephadex column.
The active fraction is collected and freeze-dried to obtain a purified formaldehyde bisulfite adduct of K582M-A. This is an example of the compound of the present invention produced as in Examples 1 and 2 based on Method 1 and Method 2 above.
The physical and physiological properties of K582M-A sodium methanesulfonate are as follows. (a) Physical properties (1) Properties White amorphous acidic substance (2) Decomposition point Gradually decomposes at 202-210℃ (3) Specific optical rotation [X] 30 D = +1.2 (1% H 2 O) ( 4) Elemental analysis value C37.4% H5.6% N15.5% (5) Molecular formula C 45 H 78 N 16 O 23 S 4 Na 4 (6) Molecular weight 1400 (vapor pressure method) (7) Color reaction Miron reaction , Sakaguchi reaction, xanthoprotein reaction positive, ninhydrin (neutral) reaction, Moritschew, Thorens reaction negative (8) Constituent amino acids (measured by hydrolysis with 6N hydrochloric acid in a sealed tube at 110°C for 24 hours) Threonine 1, Tyrosine 1, ornithine 2,
Lysine 1, arginine 1, hydroxyl arginine 1 (9) Infrared absorption spectrum Figure 1 (b) Physiological properties (1) Antibacterial activity (minimum inhibitory concentration mcg/ml)
【表】
モーマ
[Table] Moma
【表】
リス
(2) 毒性 〓マウス静脈内注射でのLD50
K582M−Ａ（原料） 120〜144mg／Kg
K582M−Ａ
1100〜1500mg／Kg
メタンスルホン酸ナトリウム
上のデータより明らかなように、K582M
−Ａのメタンスルホン酸ナトリウムの静脈内
注射での毒性は、K582M−Ａのおおよそ1/1
０であり、このように低毒性であるがため、
静脈注射等臨床への用途が拡大された。
(3) 担がんラツトにおける制がん効果、担がん
マウスにおける制がん効果、生体の免疫系に
対する作用、インターフエロン・インデユー
サー様作用においても本発明のK582M−Ａ
のメタンスルホン酸ナトリウムは原料である
K582M−Ａと同等の効果を示した。
以下に実施例を挙げて本発明の化合物の製造例
を詳説するが、これらの実施例は単なる例示であ
つて、これにより本発明が限定されるものではな
く、本発明の精神を逸脱しない改変あるいは改良
も本発明の範囲に包含されるべきである。
実施例 １
K582M−Ａ（塩酸塩）１ｇを水10mlにとか
し、これに36％ホルマリン溶液0.5ml加え、PHを
1N−NaOHでPH8.5に調整し、30分室温で撹拌す
る。生じた白色沈殿を別採取し、水洗後、水10
mlに懸濁させ、40℃の水浴中で10％重亜硫酸ナト
リウム６mlを撹拌しながら徐々に加え、２時間反
応させる。反応後この反応液をセフアデツクスＧ
−10カラム（250ml）に通し、水で溶出させ脱塩
する。脱塩後凍結乾燥するとK582M−Ａのメタ
ンスルホン酸ナトリウムが白色粉末として890mg
得られる。
実施例 ２
重亜硫酸ナトリウム１ｇを水10mlにとかし、こ
れに36％ホルマリン３mlを30℃にて撹拌しながら
加え、ホルムアルデヒド重亜硫酸ナトリウム付加
体を作る。この溶液３mlにK582M−Ａ（遊離塩
基）１ｇを加え、40℃で撹拌しながら２時間反応
させる。反応後この反応液をセフアデツクスＧ−
10カラム（250ml）に通し、水で溶出させ脱塩す
る。脱塩後、凍結乾燥すると、K582M−Ａのメ
タンスルホン酸ナトリウムが白色粉末として742
mg得られる。[Table] Squirrel
(2) Toxicity = LD in mouse intravenous injection 50 K582M-A (raw material) 120-144 mg/Kg K582M-A 1100-1500 mg/Kg Sodium methanesulfonate As is clear from the above data, K582M
-The toxicity of sodium methanesulfonate after intravenous injection of A is approximately 1/1 that of K582M-A.
0, and because of its low toxicity,
Clinical applications such as intravenous injection have been expanded. (3) The K582M-A of the present invention also has anticancer effects in tumor-bearing rats, anticancer effects in tumor-bearing mice, effects on the body's immune system, and interferon inducer-like effects.
Sodium methanesulfonate is the raw material
It showed the same effect as K582M-A. The production examples of the compounds of the present invention will be explained in detail by giving examples below, but these examples are merely illustrative and the present invention is not limited thereby, and modifications that do not depart from the spirit of the present invention may be made. Alternatively, improvements should also be included within the scope of the present invention. Example 1 Dissolve 1 g of K582M-A (hydrochloride) in 10 ml of water, add 0.5 ml of 36% formalin solution, and adjust the pH.
Adjust the pH to 8.5 with 1N NaOH and stir at room temperature for 30 minutes. Separately collect the white precipitate that has formed, wash it with water, and add 10% of water.
ml, and gradually add 6 ml of 10% sodium bisulfite with stirring in a water bath at 40°C, and react for 2 hours. After the reaction, transfer the reaction solution to Cephadex G.
Pass through a −10 column (250 ml) and desalt by eluting with water. When desalted and freeze-dried, K582M-A sodium methanesulfonate becomes 890 mg as a white powder.
can get. Example 2 1 g of sodium bisulfite is dissolved in 10 ml of water, and 3 ml of 36% formalin is added thereto with stirring at 30°C to prepare a formaldehyde sodium bisulfite adduct. Add 1 g of K582M-A (free base) to 3 ml of this solution, and react at 40° C. for 2 hours with stirring. After the reaction, the reaction solution was transferred to Sephadex G-
10 columns (250 ml) and eluted with water to desalt. After desalting and freeze-drying, K582M-A sodium methanesulfonate becomes 742 as a white powder.
mg obtained.
第１図は、本発明の目的物の１つである
K582M−Ａのメタンスルホン酸ナトリウムの赤
外線吸収スペクトルを示すものである。
FIG. 1 is one of the objects of the present invention.
This figure shows the infrared absorption spectrum of sodium methanesulfonate K582M-A.
Claims (1)
チロシン１、オルニチン２、ヒドロキシアルギニ
ン１、リジン１を構成アミノ酸とする直鎖状ペプ
チド性抗生物質である、アルギニン−γ−ヒドロ
キシアルギニン−オルニチン−スレオニン−オル
ニチン−リジンで示されるアミノ酸配列を有する
ポリペプチド（K582M−Ａ）から４個の遊離ア
ミノ基を除いた母核を示し、Ｍはアルカリ金属又
はアンモニウムイオンを示す。） で示される、アルギニン−γ−ヒドロキシアルギ
ニン−オルニチン−スレオニン−オルニチン−リ
ジンで示されるアミノ酸配列を有するポリペプチ
ド（K582M−Ａ）のホルムアルデヒド重亜硫酸
塩付加物。 ２ 一般式 Ｘ（−NH2）4 （式中、Ｘはアルギニン１、スレオニン１、チ
ロシン１、オルニチン２、ヒドロキシアルギニン
１、リジン１を構成アミノ酸とする直鎖状ペプチ
ド性抗生物質である、アルギニン−γ−ヒドロキ
シアルギニン−オルニチン−スレオニン−オルニ
チン−リジンで示されるアミノ酸配列を有するポ
リペプチド（K582M−Ａ）から４個の遊離アミ
ノ基を除いた母核を示す。） で示されるアルギニン−γ−ヒドロキシアルギニ
ン−オルニチン−スレオニン−オルニチン−リジ
ンで示されるアミノ酸配列を有するポリペプチド
（K582M−Ａ）に４倍モル量のホルムアルデヒド
を反応させ、得られたシツフ塩基に４倍モル量の
重亜硫酸塩と反応させるか又はK582M−Ａに予
めホルムアルデヒドと重亜硫酸塩との反応により
得られた４倍モル量のホルムアルデヒドの重亜硫
酸塩付加体を反応させることを特徴とする、一般
式 Ｘ（−NH−CH2−SO3M）4 （式中、Ｘは上で定義した通りであり、Ｍはア
ルカリ金属又はアンモニウムイオンを示す。） で示される、アルギニン−γ−ヒドロキシアルギ
ニン−オルニチン−スレオニン−オルニチン−リ
ジンで示されるアミノ酸配列を有するポリペプチ
ド（K582M−Ａ）のホルムアルデヒド重亜硫酸
塩付加物の製造方法。[Claims] 1 General formula X(-NH-CH 2 -SO 3 M) 4 (wherein,
A polypeptide having the amino acid sequence arginine-γ-hydroxyarginine-ornithine-threonine-ornithine-lysine, which is a linear peptide antibiotic whose constituent amino acids are 1 tyrosine, 2 ornithine, 1 hydroxyarginine, and 1 lysine. This shows the mother nucleus obtained by removing four free amino groups from (K582M-A), and M represents an alkali metal or ammonium ion. ) A formaldehyde bisulfite adduct of a polypeptide (K582M-A) having the amino acid sequence arginine-γ-hydroxyarginine-ornithine-threonine-ornithine-lysine. 2 General formula X(-NH 2 ) 4 (wherein, Arginine-γ- represented by -γ-Hydroxyarginine-Ornithine-Threonine-Ornithine-Lysine from which four free amino groups have been removed from the polypeptide (K582M-A) A polypeptide (K582M-A) having an amino acid sequence represented by hydroxyarginine-ornithine-threonine-ornithine-lysine was reacted with 4 times the molar amount of formaldehyde, and the resulting Schiff base was reacted with 4 times the molar amount of bisulfite. The general formula X(-NH-CH 2 -SO 3 M) 4 (wherein X is as defined above and M represents an alkali metal or ammonium ion) arginine-γ-hydroxyarginine-ornithine-threonine-ornithine-lysine A method for producing a formaldehyde bisulfite adduct of a polypeptide (K582M-A) having the amino acid sequence shown below.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP11178078A JPS5547697A (en) | 1978-09-13 | 1978-09-13 | Aldehyde bisulfite adduct of peptide antibiotic k582m-a and its preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP11178078A JPS5547697A (en) | 1978-09-13 | 1978-09-13 | Aldehyde bisulfite adduct of peptide antibiotic k582m-a and its preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5547697A JPS5547697A (en) | 1980-04-04 |
| JPS6218559B2 true JPS6218559B2 (en) | 1987-04-23 |
Family
ID=14569969
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP11178078A Granted JPS5547697A (en) | 1978-09-13 | 1978-09-13 | Aldehyde bisulfite adduct of peptide antibiotic k582m-a and its preparation |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5547697A (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5811812A (en) * | 1981-07-16 | 1983-01-22 | Yokogawa Hokushin Electric Corp | Memory type recorder |
| US4507740A (en) * | 1981-09-08 | 1985-03-26 | Grumman Aerospace Corporation | Programmable signal analyzer |
| JPS6240538U (en) * | 1985-08-30 | 1987-03-11 | ||
| JP4875415B2 (en) * | 2006-06-27 | 2012-02-15 | 有限会社清田製作所 | Contact probe |
-
1978
- 1978-09-13 JP JP11178078A patent/JPS5547697A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5547697A (en) | 1980-04-04 |
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