JPS5965058A - Heneicosapeptide - Google Patents
HeneicosapeptideInfo
- Publication number
- JPS5965058A JPS5965058A JP57175187A JP17518782A JPS5965058A JP S5965058 A JPS5965058 A JP S5965058A JP 57175187 A JP57175187 A JP 57175187A JP 17518782 A JP17518782 A JP 17518782A JP S5965058 A JPS5965058 A JP S5965058A
- Authority
- JP
- Japan
- Prior art keywords
- added
- solution
- gly
- stirring
- methanol
- 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.)
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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
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- Peptides Or Proteins (AREA)
Abstract
Description
【発明の詳細な説明】
いたインスリン様成長因子一l(以下,IGF−1と略
記する)の測定に関するものである。DETAILED DESCRIPTION OF THE INVENTION This invention relates to the measurement of insulin-like growth factor-1 (hereinafter abbreviated as IGF-1).
1GF−Iは1976年, Rinderknecht
らによりインスリン抗体により抑制されない血中インス
リン様活性物質の一つとして見出され( Proc.
Natl. kcadJci.USA.73: 28
65〜2369(197(3))+その後彼らによりヒ
ト血清より単離され.その70個のアミノ酸配列が明、
うかにされた( J.Bial. Ohem, 253
: 2769=2776(1978))o Zap
f らは、この抽出工GF−Iを用いて血清中1GF
−1をラジオイムノアッセイで測定し,臨床的有用性に
ついても報告した( Acta Endocrinol
、、 9 5 : 5 0 5 〜517(1980
))が、抽出法では型部精製が煩雑困難で収量も極めて
少なく,満足な方法とは言えない。1GF-I in 1976, Rinderknecht
It was discovered as one of the insulin-like active substances in the blood that is not suppressed by insulin antibodies (Proc.
Natl. kcadJci. USA. 73: 28
65-2369 (197(3)) + later isolated from human serum by them. The 70 amino acid sequence is clear.
(J. Bial. Ohem, 253
: 2769=2776 (1978)) o Zap
f et al. used this extractor GF-I to increase 1GF in serum.
-1 was measured by radioimmunoassay, and the clinical usefulness was also reported (Acta Endocrinol
, 95: 505-517 (1980
)) However, in the extraction method, purification of the mold part is complicated and difficult, and the yield is extremely low, so it cannot be said to be a satisfactory method.
その後, Zapfらにより前記抽出工GF−工を用い
たラジオイムノアッセイにより下垂体疾患等における血
清9工Gv − 工測定の意砂が確認された( J.O
lin. Invest.、 6 8 :13 21〜
l38 0(1981))が日常検査に応用し得る測定
系の確立には至っていない。Subsequently, Zapf et al. confirmed the significance of measuring serum Gv-1 in pituitary diseases etc. by radioimmunoassay using the above-mentioned extractant GF-4 (J.O.
lin. Invest. , 6 8:13 21~
1380 (1981)) has not yet established a measurement system that can be applied to routine testing.
そこで本発明者は工GF一工の優れたラジオイムノア,
セイ法を確立すべく研究を重ね。Therefore, the inventor of the present invention has developed an excellent radio immuno
Repeated research to establish the Sei method.
、工9y一工の26位から46位に相当するアミノ酸配
列(I)を有するヘンエイコサペプチドを製し,本物質
を用いた工G, F一工のラジオイムノアッセイを確立
した。We prepared a heneicosapeptide having the amino acid sequence (I) corresponding to positions 26 to 46 of G, G, F, and G, and established a radioimmunoassay of G and F, using this substance.
Aon −Lyn −Pro −Thr −Gly −
Tyr −Guy −Ser −5er−Ser −A
rg −Arg−Ala −Pro −Gin −Th
r −Gly−工1e−Val −Asp −Glu
(1)なお1本明細書において各アミノ酸残基は原則
として英語名最初の3文字で示し9例外としてABnは
アスパラギニル、 Ginはグルタミニル。Aon -Lyn -Pro -Thr -Gly -
Tyr-Guy-Ser-5er-Ser-A
rg-Arg-Ala-Pro-Gin-Th
r -Gly-ENG1e-Val -Asp -Glu
(1) In this specification, each amino acid residue is generally indicated by the first three letters of its English name.9 Exceptions are ABn for asparaginyl and Gin for glutaminyl.
工1eはイソロイシルを示す。1e indicates isoleucyl.
以下特別に示す以外は本明細書中ではこの式(1)のア
ミノ酸配列を有するペプチドをヘンエイコサペプチドと
称す。Unless otherwise specified below, the peptide having the amino acid sequence of formula (1) is referred to herein as a heneicosapeptide.
このペプチドは実施例に示した如く、適切な保護基と能
率的なフラグメンテーションにより簡便に製造すること
ができる。すなわち、 ’Ihr−Guy−工1e −
Val −Asp −Gluで表わされるアミノ酸配列
を有するヘキサペプチドのC端を保護し。This peptide can be easily produced using appropriate protecting groups and efficient fragmentation, as shown in the Examples. i.e. 'Ihr-Guy-E -
The C-terminus of a hexapeptide having an amino acid sequence represented by Val-Asp-Glu was protected.
このN端にArg −Arg −Ala −Pro −
Gin、Tyr −Gly −8er −Ser −S
er及びAsn −IIye −Pro −Thr −
Glyで表わされるアミノ酸配列を有する3種のペンタ
ペプチドを段階的に縮合させる。その後必要に応じ保護
基を脱離させる。Arg −Arg −Ala −Pro −
Gin,Tyr-Gly-8er-Ser-S
er and Asn -IIye -Pro -Thr -
Three types of pentapeptides having the amino acid sequence represented by Gly are condensed stepwise. Thereafter, the protecting group is removed if necessary.
このものは式(1)で示した基本的なアミノ酸配列を有
していれば目的に応じ多少の修飾を加えてもよく、アッ
セイ用または合成中間体として使用することができる。As long as this product has the basic amino acid sequence shown in formula (1), it may be modified to some extent depending on the purpose, and can be used for assay purposes or as a synthetic intermediate.
タトえば、ヘンエイコサペプチドから工GF−工のラジ
オイムノア、セイに必要な抗血清及びトレーサーを製造
することができる。抗血清作成のための免疫用抗原とし
てはヘンエイコサペプチドとアルブミン、グロブリン、
アスカリス抽出物等のタンパクとの縮合体を使用するの
が適当であり、トレーサーはハンターとグリーンウッド
の方法(Nature 、 194 : 494 (1
962) )を用いることにより得ることができる。In other words, the antiserum and tracer necessary for the radioimmunoassay of GF-GF can be produced from the heneicosapeptide. Antigens for immunization to create antiserum include heneicosapeptide, albumin, globulin,
It is appropriate to use a condensate with a protein such as Ascaris extract, and the tracer can be prepared using the method of Hunter and Greenwood (Nature, 194: 494 (1).
962) ).
次に実施例を挙げて説明するが、アミノ酸はグリシンを
除いてL−型を用いた。また9本明細書中に記載の記号
は次の意味を有する。Next, an example will be described, in which L-type amino acids were used except for glycine. Further, the symbols described in this specification have the following meanings.
2: ベンジルオキシカルボニル
Boa e三級ブチルオキシカルボニルToa:p−ト
ルエンスルホニル
NO7:ニトロ
0But :三級エチルエステル
OMO:メチルエステル
O8u:N−ヒドロキシコハク酸イミドエステル
。2: Benzyloxycarbonyl Boa eTertiary butyloxycarbonyl Toa: p-Toluenesulfonyl NO7: Nitro0But: Tertiary ethyl ester OMO: Methyl ester O8u: N-hydroxysuccinimide ester
.
DMF ニジメチルホルムアミド
110Bt: 1−ヒドロキシベンゾトリアゾールD0
0= ジシクロへキシルカルボジイミドTHF: テト
ラヒドロフラン
また、薄層クロマトグラフィーはシリカゲル薄層を用い
、 Rfはl−ブタノール・酢酸・水(4:1:5)を
、 Rf”は1−ブタノール・ピリジン・酢酸・水(8
0:20:6:24)を展開溶媒としたときの値である
。DMF Nidimethylformamide 110Bt: 1-Hydroxybenzotriazole D0
0 = Dicyclohexylcarbodiimide THF: Tetrahydrofuran Also, thin layer chromatography uses a thin layer of silica gel, Rf is 1-butanol/acetic acid/water (4:1:5), Rf'' is 1-butanol/pyridine/acetic acid・Water (8
0:20:6:24) as the developing solvent.
例1:原料の製法
A、 (1) Z −Thr−Gly−OMeZ −
Thr−OH10,139をDMF75−に溶かした溶
液に、 HOBt 5.416水溶性カルボジイミド塩
酸塩7.669及びH−Guy −OMe塩酸塩5.2
89のトリエチルアミン(5,6,7)を含むDMF(
75m+/)溶液を0℃で攪拌しながら添加した。混合
物を0℃で2時間、さ
らに室温で24時間攪拌したのち溶媒
を留去した。Example 1: Raw material manufacturing method A, (1) Z -Thr-Gly-OMeZ -
In a solution of Thr-OH10,139 in DMF75-, HOBt 5.416 water-soluble carbodiimide hydrochloride 7.669 and H-Guy-OMe hydrochloride 5.2
DMF containing 89 triethylamine (5,6,7) (
75m+/) solution was added with stirring at 0°C. The mixture was stirred at 0° C. for 2 hours and then at room temperature for 24 hours, and then the solvent was distilled off.
残渣を酢酸エチルに溶解し、IN− クエン酸、飽和炭酸水素ナトリウム溶 液及び飽和食塩水で洗浄したのち、無 水硫酸ナトリウム上で乾燥し、ついで 溶媒を留去した(以上の処理操作を以 下常法と称す。)。The residue was dissolved in ethyl acetate and IN- Citric acid, saturated sodium bicarbonate solution After washing with liquid and saturated saline, Dry over sodium hydrosulfate, then The solvent was distilled off (the above process was repeated as follows). It is called the lower common law. ).
残液をn−ヘキサンで処理すること
により固化させたのち、酢酸エチル・
n−ヘキサンより再結晶した。収量
10.839.融点115〜116℃0Rf’ 0.6
? 、 Rf 0.72゜〔α)、%9−65.0
°(0−1,07,メタノール)。The residual liquid was solidified by treatment with n-hexane, and then recrystallized from ethyl acetate/n-hexane. Yield 10.839. Melting point 115-116℃0Rf' 0.6
? , Rf 0.72゜[α), %9-65.0
°(0-1,07, methanol).
元素分析 C,。H,N、O,として
計算値 057.00. H6,46,N 9.97
実験値 056.96. H6,24,N 9.90
(2) Z −Pro −Thr −Guy −OM
e上記生成物5.3359をIN−塩酸
を含むメタノール(16,45,ag)中でパラジウム
黒を触媒として17時間接
接触光を行なった。濾過後溶媒を留去
し、残渣を五酸化リン・水酸化カリウ
ム上で乾燥した( Rf’ 0.36 、 Rf”o、
7o)。Elemental analysis C. Calculated value as H, N, O, 057.00. H6, 46, N 9.97
Experimental value 056.96. H6, 24, N 9.90
(2) Z -Pro -Thr -Guy -OM
e The above product 5.3359 was exposed to contact light for 17 hours in methanol (16,45, ag) containing IN-hydrochloric acid using palladium black as a catalyst. After filtration, the solvent was distilled off, and the residue was dried over phosphorus pentoxide/potassium hydroxide (Rf' 0.36, Rf''o,
7o).
Z −Pro−0H8,699のDMF(15m)溶液
にN−メチルモルホリ
ン1.63−及びクロルギ酸イソブチル1.92−を−
15℃で攪拌しながら加えた。これに上記還元体のトリ
エチル
アミン(2,31mg)を含むDMIF(15m)溶液
を一り5℃°で攪拌しながら加えたのち一10℃で1分
間攪拌
した。この混合物を濃縮したのち酢酸
エチルに溶解し常法により処理した。In a DMF (15 m) solution of Z-Pro-0H8,699, 1.63- of N-methylmorpholine and 1.92- of isobutyl chloroformate were added.
It was added with stirring at 15°C. To this was added a solution of DMIF (15 m) containing the above reduced triethylamine (2.31 mg) at 5° C. with stirring, followed by stirring at -10° C. for 1 minute. After concentrating this mixture, it was dissolved in ethyl acetate and treated in a conventional manner.
得られた油状の残渣に石油エーテルを
加え固化させたのち、酢酸エチル・n
−ヘキサンから再結晶して目的のトリ
ペプチドを得た。収ff14.13り、融点元素分析
02゜H27N307とじて計算値 057.00.
H6,46,N 9.97実験値 056.96.
H6,24,N 9.90(3) Z −Iys
(ToB) −Pro−Thr−Gly−OMeZ −
Pro−Thr−Gly−OMe 4.159をメタノ
ール中でパラジウム黒を触媒と
して20時間接接触光した。濾過後。Petroleum ether was added to the obtained oily residue to solidify it, and then recrystallized from ethyl acetate/n-hexane to obtain the desired tripeptide. Convergence ff14.13, melting point elemental analysis
Calculated value as 02°H27N307 057.00.
H6,46,N 9.97 Experimental value 056.96.
H6,24,N 9.90(3) Z-Iys
(ToB) -Pro-Thr-Gly-OMeZ -
Pro-Thr-Gly-OMe 4.159 was contacted with light for 20 hours in methanol using palladium black as a catalyst. After filtration.
溶媒を留去し残液をエーテルで処理し たのち減圧乾燥し、固化物を得た。The solvent was distilled off and the residual liquid was treated with ether. Afterwards, it was dried under reduced pressure to obtain a solidified product.
Z −Lys (Toe) −OH4,289をDMF
50−に溶かした溶液にHOBtL609.DOO2,
089及び上記
還元体のDMIF (50sd)溶液を0℃で攪拌しな
がら加えた。混合物を0℃
で2時間、さらに室温で20時間攪拌
したのち濃縮し、酢酸エチルに溶解し
た。これを常法により処理し、得られ
た油状の残渣にn−ヘキサンを加えガ
ム状の物質を得た。このガム状の物質。Z -Lys (Toe) -OH4,289 in DMF
HOBtL609. DOO2,
A solution of 089 and the above reduced product in DMIF (50 sd) was added at 0° C. with stirring. The mixture was stirred at 0° C. for 2 hours and then at room temperature for 20 hours, concentrated, and dissolved in ethyl acetate. This was treated in a conventional manner, and n-hexane was added to the resulting oily residue to obtain a gummy substance. This gummy substance.
をエーテルで処理したのち減圧乾燥し
固化物を得た。収量5.9(1゜
Rf O,74,Rf O,83゜〔α〕腎−50
,3°(0−2,11,メタノ−ル)。was treated with ether and then dried under reduced pressure to obtain a solidified product. Yield 5.9 (1°Rf O, 74, Rf O, 83° [α] Kidney-50
, 3° (0-2, 11, methanol).
元素分析 018H45’1lolosとシテ計算値
056.82. H6,44,N 9.95実験値
056.18. H6,42,N 9.69(4)
Z−Asn−IIys(Tos)−Pro−’Thr
−Gly−OMθ
上記生成物2.829をIN−塩酸を
含むメタノール(sob)中でパラジ
ウム黒を触媒として80時間接触還元
を行なった。濾過後濃縮し、残渣をエ
ーテル処理したのちエーテルをデカン
テーションにより除き、メタノール−
エーテルを加えて固化させた( Rtlo、80 、
Rrll 0.65 )。Elemental analysis 018H45'1lolos and shite calculation value
056.82. H6, 44, N 9.95 experimental value
056.18. H6,42,N 9.69(4)
Z-Asn-IIys(Tos)-Pro-'Thr
-Gly-OMθ The above product 2.829 was subjected to catalytic reduction in methanol (sob) containing IN-hydrochloric acid using palladium black as a catalyst for 80 hours. After filtration and concentration, the residue was treated with ether, the ether was removed by decantation, and methanol-ether was added to solidify (Rtlo, 80,
Rrll 0.65).
上記還元体をトリエチルアミンを含
むDMF2amに溶解した溶液に2−
Asn−osu 1.45りをot+で攪拌しながら加
えた。混合物を4℃で15時間
攪拌し、さらにz −Asn−08u O,299を0
℃で攪拌しながら加えたのち。To a solution in which the above reduced product was dissolved in DMF 2am containing triethylamine, 1.45 μl of 2-Asn-osu was added while stirring at ot+. The mixture was stirred at 4°C for 15 hours, and z-Asn-08u O,299 was added to 0.
After adding with stirring at ℃.
4℃で5時間攪拌した。溶媒を留去し たのち、残渣を0℃でIN−クエン酸 で処理した。得られたガム状物に水を 加え、さらに酢酸エチルを加えて懸濁 させた。有機層を水洗したのち、濃縮 乾固した。収量2.48g。融点162〜168℃。The mixture was stirred at 4°C for 5 hours. distill off the solvent Afterwards, the residue was diluted with IN-citric acid at 0°C. Processed with. Add water to the resulting gum. Add ethyl acetate and suspend. I let it happen. After washing the organic layer with water, concentrate It was dried and solidified. Yield: 2.48g. Melting point 162-168°C.
Rf’ 0.48 、 Rf O,78゜〔α)p−
2o、7°(0−1,24,DMF)元素分析 CI?
H1lIN?ol!s として計算値 054.33.
H6,28,N IIJ)9実験値 054.28
. H6,26,N 11.76「を加水分解物中の
アミノ酸の比率
Asp O,95、G171.01 、 Pr。Rf' 0.48, Rf O, 78° [α) p-
2o, 7° (0-1, 24, DMF) elemental analysis CI?
H1lIN? ol! Calculated value as s 054.33.
H6,28,N IIJ)9 Experimental value 054.28
.. H6,26,N 11.76'Proportion of amino acids in the hydrolyzateAsp O,95,G171.01, Pr.
L、08 、 Thr 1.00 、 Lys O,
90(5) Z−Asn−I7ys(Tos)−Pr
o−Thr−Gly B・−NIINH2
Z −Asn −TJys (Tos ) −Pro
−Thr −Gly−OMe 1.00 pをメタノ
ール8o−に溶解し、0℃でヒドラジン水和物
0.6−を加えた。混合物を室温で2日間放置し、濃縮
し1−ブタノールに溶
解したのち水で洗浄した。溶媒を留去
したのち残渣にエーテルを加え固化さ
せた。メタノール・エーテルよす再結
晶を行ない製品とした。収量0.88り。L, 08, Thr 1.00, Lys O,
90(5) Z-Asn-I7ys(Tos)-Pr
o-Thr-Gly B・-NIINH2 Z -Asn -TJys (Tos) -Pro
1.00 p of -Thr -Gly-OMe was dissolved in 8 o of methanol, and 0.6 o of hydrazine hydrate was added at 0°C. The mixture was left at room temperature for 2 days, concentrated, dissolved in 1-butanol, and washed with water. After the solvent was distilled off, ether was added to the residue to solidify it. The product was obtained by recrystallizing with methanol and ether. Yield: 0.88 ri.
融点173〜174℃。Melting point: 173-174°C.
Rf’ 0.40 、 Rf O,74゜元素分析
’116H111’1lolls ’ +Htoとして
計算値 052.29. H6,L4. N 15
.24実験値 052゜88. H6,21,M 1
4.99酸加水分解物中のアミノ酸の比率
Asp 1.08 、 Gly 1.01 、 Pr。Rf' 0.40, Rf O, 74° elemental analysis
Calculated value as '116H111'1lolls' +Hto 052.29. H6, L4. N15
.. 24 Experimental value 052°88. H6, 21, M 1
4.99Ratio of amino acids in acid hydrolyzate Asp 1.08 , Gly 1.01 , Pr.
O,99、Thr O,92、Lys O,9,1(1
) −Boa −Tyr −Gly −NHNH2B
oa−Tyr−OH5,649をDMIF40%tに溶
かした溶液にHOBt 2.709゜DOO4,12g
及びH−Gly −OMe塩酸塩8.009のトリエチ
ルアミン
(3,86,7)を含むDMF(20r/)溶液を0℃
で攪拌しながら添加した。O,99, Thr O,92, Lys O,9,1(1
) -Boa -Tyr -Gly -NHNH2B
Add HOBt 2.709°DOO4,12g to a solution of oa-Tyr-OH5,649 dissolved in DMIF40%t.
A DMF (20 r/) solution containing triethylamine (3,86,7) and H-Gly-OMe hydrochloride 8.009 was heated at 0°C.
was added while stirring.
混合物を0℃で1時間さらに室温で 20時間攪拌したのち溶媒を留去し酢 酸エチルに溶解した。これを常法にて 処理し、得られた油状物にエーテル・ n−ヘキサンを加え結晶化させたのち メタノール−エーテル(1:1)に溶 解し、活性炭処理のあと、酢酸エチル ・n−ヘキサンから再結晶を行ない。The mixture was heated at 0°C for 1 hour and then at room temperature. After stirring for 20 hours, the solvent was distilled off and vinegar was added. Dissolved in ethyl acid. Do this in the usual way The resulting oil is treated with ether and After adding n-hexane and crystallizing Dissolved in methanol-ether (1:1) After decomposition and activated carbon treatment, ethyl acetate - Recrystallize from n-hexane.
ジペプチドのメチルエステル体 5.799を得た(RfO,85゜ Rf O,88)。Methyl ester of dipeptide 5.799 was obtained (RfO, 85° Rf O, 88).
上記メチルエステルをメタノール
50−に溶解し、ヒドラジン水和物
8.9+++lを加えたのち、室温で24時間放置した
。混合物を水を飽和した1−
ブタノールで抽出したのち、メタノー
ル−エーテルから再結晶を行ない目的
物を得た。収fi5.019.融点157〜160℃。The above methyl ester was dissolved in 50 methanol, 8.9 +++ l of hydrazine hydrate was added, and the mixture was left at room temperature for 24 hours. The mixture was extracted with 1-butanol saturated with water, and then recrystallized from methanol-ether to obtain the desired product. Collection fi5.019. Melting point: 157-160°C.
Rf’ 0.67 、 Rf” 0.77゜〔α)%’
+1 a、oo(0−L91. 、ilタノール〕。Rf' 0.67, Rf"0.77゜[α)%'
+1 a, oo (0-L91., iltanol).
元素分析 0.。H24N405として計算値 054
.5(、H6,87,N 15.90実験値 054.
70. H6,8−2,N 15.88(2) Z
−Ser −Sor −OMeZ−F3er−HHH
H,2,589をDMI+’20−に溶カルだ溶液に6
N−塩酸/
ジオキサン5.01−及び亜硝酸イソアミルト34−を
一15℃で激しく攪拌
しながら加えた。混合物をトリエチル
アミンで中和したのちH−Ser −OMel、859
のトリエチルアミン
(1,68,j)を含むDMlr (20常l)溶液を
一15℃で加えた。混合物を
一10℃で2時間、さらに4℃で20
時間攪拌したのち溶媒を留去し酢薗エ
チルに溶解した。これを常法にて処理
したのちエーテルを加えて結晶化させ。Elemental analysis 0. . Calculated value as H24N405 054
.. 5 (, H6, 87, N 15.90 experimental value 054.
70. H6,8-2,N 15.88(2) Z
-Ser -Sor -OMeZ-F3er-HHH
Dissolve H, 2,589 in DMI+'20- and add 6
N-hydrochloric acid/dioxane 5.01 and isoamyl nitrite 34 were added at -15° C. with vigorous stirring. After neutralizing the mixture with triethylamine, H-Ser-OMel, 859
A solution of DMlr (20 liters) containing triethylamine (1,68,j) was added at -15°C. The mixture was stirred at -10°C for 2 hours and then at 4°C for 20 hours, then the solvent was distilled off and the mixture was dissolved in ethyl vinegar. After treating this in a conventional manner, ether was added to crystallize it.
サラにメタノール−エーテルから再結
晶して製品とした。収512.529゜融点142〜1
43℃。The product was then recrystallized from methanol-ether. Yield 512.529° Melting point 142-1
43℃.
〔α〕背−4,2°(0−1,14,メタノール)元素
分析 CヤH2ON2O?として
計算値 052.94. H5,92,N 8.1?
実験値 05&13. H5,92,N 8.17(
8) Z 7 Ser −Ser −Ser −OM
。[α] Back -4,2° (0-1,14, methanol) Elemental analysis Cya H2ON2O? Calculated value as 052.94. H5, 92, N 8.1?
Experimental value 05 & 13. H5, 92, N 8.17 (
8) Z7 Ser -Ser -Ser -OM
.
Z−3er−8er−OMe 4.709をIN−塩酸
を含むメタノール(1,3,8m)中でパラジウム黒を
触媒として26時間
接触還元を行なった。濾過後溶媒を留
失して固化物を得た。Z-3er-8er-OMe 4.709 was subjected to catalytic reduction for 26 hours in methanol (1,3,8m) containing IN-hydrochloric acid using palladium black as a catalyst. After filtration, the solvent was distilled off to obtain a solidified product.
Z−8er−NHNH24,199の6N−塩酸/ジオ
キサン(8,30td)を含むDMy(3og)溶液に
一15℃で攪
拌しながら亜硝酸イソアミル2.22dを加え、トリエ
チルアミンで中和した
のち、上記還元体のトリエチルアミン
(1,93,j)を含むDMF (80gLt)溶液を
一15℃で攪拌しながら加えた。To a DMy (3 og) solution containing Z-8er-NHNH24,199 in 6N-hydrochloric acid/dioxane (8,30 td), 2.22 d of isoamyl nitrite was added while stirring at -15°C, and after neutralization with triethylamine, the above A DMF (80 gLt) solution containing the reduced product triethylamine (1,93,j) was added at -15° C. with stirring.
混合物を一15℃で1時間、さらに 4℃で20時間攪拌したのち溶媒を留 失し、残渣を水で飽和した1−ブタノ ールで抽出した。濃縮し、酢酸エチル から結晶化させ、さらにメタノール・ 酢酸エチルから再結晶し製品とした。Heat the mixture at -15°C for 1 hour, then After stirring at 4℃ for 20 hours, the solvent was distilled off. 1-butano and the residue was saturated with water. Extracted with a tool. Concentrate to ethyl acetate Crystallized from methanol and The product was recrystallized from ethyl acetate.
収量4,159 、融点204〜207℃。Yield 4,159, melting point 204-207°C.
Rfl 0.67、Rfl 0.76 。Rfl 0.67 , Rfl 0.76 .
〔α) ”+ 10.7°(0−1,18,DMF )
元素分析 C111H2sN80oとして計算値 05
0.58. H5,90,N 9.83実験値 05
0.39. H5,76、N 9.79(4) B
oC−Tyr −Gly −Ser −Ser −So
r −OMeZ−8er−8er−8er−OMe
2.059を10%酢酸(20m)を含むメタノー
ル(150at )中でパラジウム黒を触媒として15
時間接触還元を行なった。[α)”+10.7°(0-1,18,DMF)
Elemental analysis Calculated value as C111H2sN80o 05
0.58. H5, 90, N 9.83 experimental value 05
0.39. H5,76, N 9.79 (4) B
oC-Tyr-Gly-Ser-Ser-So
r -OMeZ-8er-8er-8er-OMe
2.059 was dissolved in methanol (150 at) containing 10% acetic acid (20 m) using palladium black as a catalyst.
Time contact reduction was performed.
濾過後、溶媒を留去し残渣をエーテル
により固化した( Rrl O,19、RtRO,57
)。After filtration, the solvent was distilled off and the residue was solidified with ether (RrlO, 19, RtRO, 57
).
Boo −Tyr −Gly −NHNH22,039
をDMFIO艷に溶かした溶液に6N−
塩酸/ジオキサン2.884を一15℃で攪拌しなから
°加えたのち、亜硝酸イソアミル0.77−を加え、ト
リエチルアミンで中和した。この溶液に上記還
元体のトリエチルアミン(0,67,7)を含むDMF
(10コ)溶液を一15℃で攪拌しながら加え、−1
5℃で1時
間、さらに4℃で15時間攪拌した。Boo-Tyr-Gly-NHNH22,039
To a solution prepared by dissolving 6N hydrochloric acid/dioxane in a DMFIO vessel, 2.884 g of 6N hydrochloric acid/dioxane was added at -15° C. while stirring, and then 0.77 g of isoamyl nitrite was added, followed by neutralization with triethylamine. DMF containing the above reduced triethylamine (0,67,7) in this solution
(10) Add the solution while stirring at -15℃, -1
The mixture was stirred at 5°C for 1 hour and then at 4°C for 15 hours.
溶媒を留去し、残渣を水に溶解し酢酸 でpHを8−4に調整したのち、1− ブタノールで抽出した。溶媒を留去し たのち残渣をエーテルにより固化し。The solvent was distilled off, the residue was dissolved in water, and acetic acid was added. After adjusting the pH to 8-4 with Extracted with butanol. distill off the solvent Afterwards, the residue was solidified with ether.
さらにメタノール・エーテルから再結
晶し製品とした。収量2.549.融点156〜158
℃。The product was further recrystallized from methanol/ether. Yield 2.549. Melting point 156-158
℃.
〔α麿−13,2°(0−1,00,メタノール〕。
C6
元素分析 C工H8゜N、O□、・−)H2Oとして計
算値 050.40. H6,45,N 11.80
実験値 050.35. H6,35,H11,28
(5) Boa −Tyr −Gly = Ser
−Sor −Ser −NHNH。[αMaro-13,2° (0-1,00, methanol)].
C6 Elemental analysis C engineering H8°N, O□, ·-) Calculated value as H2O 050.40. H6,45,N 11.80
Experimental value 050.35. H6, 35, H11, 28
(5) Boa −Tyr −Gly = Ser
-Sor -Ser -NHNH.
BQO−Tyr −Guy −Ser −5er−タe
r −OMel、22gをメタノール・エタノール
(1:1)20−に溶かした溶液に
0℃でビドラジン水和物1.0−を加えた。混合物を室
温で20時間放置した
のち、沈澱物を濾別し、エタノールで
洗浄した。得られた粗生成物1.229をメタノールに
懸濁させ、エーテルに
より沈澱させ目的物を得た。収量
1.1 a 9 、融点215℃(分解)。BQO-Tyr-Guy-Ser-5er-tae
1.0-hydrazine hydrate was added to a solution of 22 g of r-OMel dissolved in 20-methanol/ethanol (1:1) at 0°C. After the mixture was left at room temperature for 20 hours, the precipitate was filtered off and washed with ethanol. The obtained crude product (1.229 g) was suspended in methanol and precipitated with ether to obtain the desired product. Yield 1.1 a 9 , melting point 215°C (decomposed).
Rfl O,4B 、 Rfl 0.73゜〔α〕曾−
83.6°(0−1,04,水)。Rfl O,4B, Rfl 0.73゜[α] So-
83.6° (0-1,04, water).
元素分析 ’!1lH1lN?011として計算値 0
48.93. H6,41,N 15.9B実験値
048.87. H6,58,N 15.53(1)
Z −Pro−Gin −NHmHBocZ−Gi
n−11HIJHBoc 7.899をメタノール中で
パラジウム黒を触媒として接
触還元した。濾過し、溶媒を留去した
のち、残渣をエーテルで処理した(Rflo、47.R
fl 0.64) 。Elemental analysis'! 1lH1lN? Calculated value as 011 0
48.93. H6,41,N 15.9B experimental value
048.87. H6,58,N 15.53(1)
Z-Pro-Gin-NHmHBocZ-Gi
n-11HIJHBoc 7.899 was catalytically reduced in methanol using palladium black as a catalyst. After filtration and evaporation of the solvent, the residue was treated with ether (Rflo, 47.R
fl 0.64).
Z−Pro−OH4,499をDMP15mに溶解した
溶液にN−メチルモルホリ
ブ′
ン1.98−及びクロルギ酸イソ←チル2.84−を−
18℃で攪拌しながら加えた。これに上記還元体のN−
メチル
モルホリン(g、2o−)を含むDMF(10m)溶液
を−18℃で攪拌しな
がら加え一10℃で1分間攪拌した。In a solution of Z-Pro-OH4,499 dissolved in DMP15m, N-methylmorpholibone 1.98- and iso←thyl chloroformate 2.84- were added.
The mixture was added at 18° C. with stirring. To this, the N-
A DMF (10 m) solution containing methylmorpholine (g, 2o-) was added at -18°C with stirring, and the mixture was stirred at -10°C for 1 minute.
溶媒を留去したのち、酢酸エチルに溶 解し常法により処理した。残渣をエー テルによす固化し、メタノール・エー テルから再結晶して製品とした。収量 ?、619 、融点177〜179℃。After distilling off the solvent, dissolve in ethyl acetate. It was then treated in a conventional manner. Remove the residue It solidifies into methanol and methanol. The product was recrystallized from tel. yield ? , 619, melting point 177-179°C.
Rfl O,77、Rfl O,86゜(α)D−71
8°(0−2,55,メタノール)
元素分析 ’tgHssNso’r として計算値 0
56.26. H6,’l’l、 N 14.25
実験値 056.19. H6,80,N 14.2
9(2) Z −Ala −Pro −Gin −N
HIJHBoaZ −Pro −Gln −NHHHB
oa 5.76gをメタノール中でパラジウム黒を触
媒と
して4時間接触還元した。濾過後溶媒
を留去し固化物を得た( Rfl O,85。Rfl O, 77, Rfl O, 86° (α) D-71
8° (0-2,55, methanol) Elemental analysis Calculated value as 'tgHssNso'r 0
56.26. H6,'l'l, N 14.25
Experimental value 056.19. H6,80,N 14.2
9(2) Z -Ala -Pro -Gin -N
HIJHBoaZ -Pro -Gln -NHHHB
5.76 g of OA was catalytically reduced in methanol using palladium black as a catalyst for 4 hours. After filtration, the solvent was distilled off to obtain a solidified product (Rfl O, 85).
Rf璽 0.65)。Rf seal 0.65).
Z−Ala−OH2,629をDMF
15−に溶解した溶液にN−メチルモ
ルホリン1.29−及びクロルギ酸イソブチル1.52
−を−18℃で撹拌しながら加えたのち、上記還元体の
N−メ
チルモルホリン(1,29m)を含む
DMF (15tg)溶液を一18℃で攪拌しながら加
え一10℃で1分間攪拌
した。溶媒を留去したのち、酢酸エチ
ルで抽出し、エーテルにより固化させ。A solution of Z-Ala-OH2,629 in DMF 15- was added with 1.29- and 1.52-N-methylmorpholine and isobutyl chloroformate.
- was added with stirring at -18°C, then a DMF (15tg) solution containing the above reduced N-methylmorpholine (1,29m) was added with stirring at -18°C, and the mixture was stirred at -10°C for 1 minute. . After distilling off the solvent, the mixture was extracted with ethyl acetate and solidified with ether.
メタノール・エーテルから再結晶した。Recrystallized from methanol/ether.
収量5.549.融点114〜117℃。Yield 5.549. Melting point 114-117°C.
Rfl O,71,Rf履 0.85゜〔α)”−8
8,4°(0−1,85,メタノール)。Rfl O, 71, Rf 0.85゜〔α)”-8
8,4° (0-1,85, methanol).
元素分析 ’26H38N60g・+H20として計算
値 054.63. H6,88,N 14.70実
験値 05.4.68. H6,89,N 14..
76(a) Z−Arg (No、 ) −Ala−
Pro−Gln−NHNHBoc
Z−Ala−Pro−Gin−NHNHBoc 2.2
59をメタノール5〇−中でパラジウム黒
を触媒として155時間接触還した。Elemental analysis '26H38N60g・+H20 Calculated value 054.63. H6,88,N 14.70 Experimental value 05.4.68. H6,89,N 14. ..
76(a) Z-Arg (No, ) -Ala-
Pro-Gln-NHNHBoc Z-Ala-Pro-Gin-NHNHBoc 2.2
59 was catalytically reduced in 50 methanol using palladium black as a catalyst for 155 hours.
濾過後溶媒を留去し、残渣にエーテル を加えたのち乾燥させ固化物を得た ( Rfl O,26、Rfl O,60)。After filtration, the solvent was distilled off and the residue was diluted with ether. was added and dried to obtain a solidified product. (Rfl O, 26, Rfl O, 60).
Z−Arg(No、)−0H1,299を6Mア10−
に溶かした溶液にN−メチルモ
ルホリン0.4−及びクロルギ酸イソブチル0.47−
を−18℃で攪拌しながら加えた。この混合物に上記還
元体の
N−メチルモルホリン(0,4m)を含むDMF(lO
−)溶液を°−15℃で攪拌しながら加えたのち一10
℃で1
分間攪拌した。溶媒を留去したのち。Z-Arg(No,)-0H1,299 to 6M A10-
N-methylmorpholine 0.4- and isobutyl chloroformate 0.47-
was added with stirring at -18°C. This mixture contains DMF (lO
-) After adding the solution with stirring at -15°C,
The mixture was stirred at ℃ for 1 minute. After distilling off the solvent.
残渣にエーテルを加えガム状物を得。Add ether to the residue to obtain a gum-like substance.
飽和炭酸水素す) IJウム溶液で洗浄し ゛1−ブタ
ノールψ酢酸エチル(1: 1゜150.11/)に溶
解した。有機層を水で洗浄したのち10%酢酸でpH4
・0に調節し、1−ブタノールで飽和した
2%酢酸溶液で洗浄した。有機層をま
とめて溶媒を留去したのち、残渣をエ
ーテルから固化し、メタノール・エー
テルより再結晶し°(目的物を得た。The mixture was washed with a saturated IJ solution and dissolved in 1-butanol/ethyl acetate (1:1/150.11/). After washing the organic layer with water, the pH was adjusted to 4 with 10% acetic acid.
- Adjusted to 0 and washed with 2% acetic acid solution saturated with 1-butanol. After the organic layers were combined and the solvent was distilled off, the residue was solidified from ether and recrystallized from methanol/ether (the desired product was obtained).
収量2,719 、融点152℃(分解)。Yield 2,719, melting point 152°C (decomposition).
Rfl O,56、Rfl O,82゜〔α〕背−39
,8°(0−1,02,DMF)元素分析 ’j2H4
9Nl1011・+H,Oとして計算値 049.73
. H6,52,N 14゜94実験値 049.6
5. H6,42,N 14.88(4) Z −
Arg (No、 ) −Arg −Ala−Pro
−Gin −NHNHBo。Rfl O, 56, Rfl O, 82° [α] back -39
,8°(0-1,02,DMF) elemental analysis 'j2H4
Calculated value as 9Nl1011・+H,O 049.73
.. H6,52,N 14°94 Experimental value 049.6
5. H6,42,N 14.88(4) Z −
Arg (No, ) -Arg -Ala-Pro
-Gin-NHNHBo.
Z −Arg (No2) −Ala −Pro −G
in −NHIJHBoc 2,78 gをメタノー
ル(50+d)、酢酸(8−)及び水
(42,g)の混合液中パラジウム黒を触媒として40
0時間接触還した。Z -Arg (No2) -Ala -Pro -G
in -NHIJHBoc 2,78 g was dissolved in a mixture of methanol (50+d), acetic acid (8-) and water (42, g) using palladium black as a catalyst.
The contact was returned for 0 hours.
濾過後、濾液を濃縮し、残渣を凍結乾
燥し、固化物を得た(urlO,17゜RflI O,
63)。After filtration, the filtrate was concentrated and the residue was freeze-dried to obtain a solidified product (urlO, 17°RflIO,
63).
Z −Arg (NO2) −OH1−169をDMF
10c+tIC溶かした溶液にN−メチルモ D。Z -Arg (NO2) -OH1-169 in DMF
Add N-methylmoD to the solution containing 10c+tIC.
ルボリン0.36−及びクロルギ酸イソブチル0.42
−を一18℃で攪拌しながら加えた。この混合物に上記
還元体
のDMF (15s+t)溶液を一18℃で攪拌しなが
ら加えたのち一10℃で1
分間攪拌した。溶媒を留去し、残渣を
酢酸エチルから固化させた。得られた
粗生成物(L2.59) を1−ブタノールに溶解し
、2%酢酸溶液で洗浄した。Ruborin 0.36- and isobutyl chloroformate 0.42-
- was added with stirring at -18°C. A DMF (15s+t) solution of the above reduced product was added to this mixture at -18°C with stirring, and then stirred at -10°C for 1 minute. The solvent was evaporated and the residue was solidified from ethyl acetate. The obtained crude product (L2.59) was dissolved in 1-butanol and washed with 2% acetic acid solution.
溶媒を留去したのち残渣をエーテルに より固化し、さらにメタノール・酢酸 エチルから再結晶を行ない製品とした。After distilling off the solvent, the residue was dissolved in ether. It becomes more solidified and further methanol/acetic acid The product was recrystallized from ethyl.
収量2.39 g、融点166〜168℃。Yield 2.39 g, melting point 166-168°C.
Rfl 0037 、 Rfl O,7?。Rfl 0037 , Rfl O,7? .
〔α〕¥)−aa、ao(0−1,15,メタノール)
元素分析 Cl1lH6jNj5012・OH,0OO
H・2H20として
計算値 048゜041. H6,95,N 2]、
、01実験値 047.85. H6,62,’N
21.02(リ Z−’lhr −Gly−NHNH
。[α] ¥)-aa, ao (0-1,15, methanol)
Elemental analysis Cl1lH6jNj5012・OH,0OO
Calculated value as H・2H20 048°041. H6,95,N2],
, 01 experimental value 047.85. H6,62,'N
21.02 (Li Z-'lhr -Gly-NHNH
.
Z−Thr−Gly−OMe 4.39のエタノール(
80m)溶液にヒドラジン水和物
フ一を0℃で加え、室温で20時間放
置した。沈澱を濾別し、濃硫酸及び五
酸化リン上で減圧乾燥した。得られた
粗生成物(4,229)を熱エタノールに懸濁し、冷却
した後、沈澱物を濾取
した。収fiL97g、融点170〜
171℃。Z-Thr-Gly-OMe 4.39 ethanol (
80m) To the solution was added hydrazine hydrate at 0°C, and the mixture was left at room temperature for 20 hours. The precipitate was filtered off and dried under reduced pressure over concentrated sulfuric acid and phosphorus pentoxide. The obtained crude product (4,229) was suspended in hot ethanol, and after cooling, the precipitate was collected by filtration. Yield: 97g, melting point: 170-171°C.
Rfl O,57、Rfl O,82゜〔α〕蟹−Ll
’ (0−1,88,D M F’ )元素分析 Cl
4H2oN405として計算値 051.85. H
6,22,N 17.27実験値 051.68.
H6,19,N 17.24(2) Z−Va
l−Aep(ODut)−Glu(OBut)2Z−A
sp(OBut) −OH6,479のTHF(50−
3溶液にN−メチルモルポリ
> 22 vd及びクロルギ酸イソブチル2.64□−
を−18℃で攪拌しながら加えた。Rfl O, 57, Rfl O, 82° [α] Crab-Ll
'(0-1,88,DM F') Elemental analysis Cl
Calculated value as 4H2oN405 051.85. H
6,22,N 17.27 Experimental value 051.68.
H6,19,N 17.24(2) Z-Va
l-Aep(ODut)-Glu(OBut)2Z-A
sp(OBut) -OH6,479 THF(50-
3 solution of N-methylmolpoly>22 vd and isobutyl chloroformate 2.64□-
was added with stirring at -18°C.
この溶液にH−Glu (0But )、塩酸塩5.9
29のN−メチルモルホリン
(2,2*/ )を含むTHF (5o@t)の溶液を
一18℃で攪拌しながら加えたのち
一10℃で1分間攪拌した。溶媒を留去したのち、残渣
をエーテルに溶解し、常法により処理した。残渣をn−
ヘキサンで2回処理したのち、減圧乾燥してジ
ペプチドZ −Asp (OBut)−Glu(OBi
zt)210.049(油状物)を得た(nf O,8
6゜
RrO,83)。得られた油状物をメ
タノール中でパラジウム黒を触媒とし
て2時間接触還元を行なった(Rf’
0.73 、 Rf O,8)。濾過し、溶媒を留去
したのち、残液を減圧乾燥した。In this solution, H-Glu (0But), hydrochloride 5.9
A solution of THF (5o@t) containing 29 N-methylmorpholine (2,2*/ ) was added with stirring at -18°C, and then stirred at -10°C for 1 minute. After evaporating the solvent, the residue was dissolved in ether and treated in a conventional manner. Residue n-
After treatment with hexane twice, the dipeptide Z-Asp(OBut)-Glu(OBi
zt) 210.049 (oil) was obtained (nf O,8
6°RrO, 83). The obtained oil was subjected to catalytic reduction in methanol using palladium black as a catalyst for 2 hours (Rf' 0.73, Rf O, 8). After filtration and distilling off the solvent, the residual liquid was dried under reduced pressure.
Z−’Val−OH4,029のT HF(30m)溶
液にN−メチルモルホリ
ン(1,76、/)及びクロルギ酸イソブチル2.11
−を−18℃で攪拌しながら加えた。この溶液に上記還
元体のN
−メチルモルホリン(1,76−d)を含むTIP(8
0gIり溶液を一18℃で攪拌しながら加えたのち、−
10℃で
1分間攪拌した。溶媒を留去したのち。A solution of Z-'Val-OH4,029 in THF (30 m) contains N-methylmorpholine (1,76,/) and isobutyl chloroformate 2.11
- was added with stirring at -18°C. This solution contains TIP (8) containing the reduced form N-methylmorpholine (1,76-d).
After adding 0gI solution with stirring at -18℃, -
The mixture was stirred at 10°C for 1 minute. After distilling off the solvent.
残渣をエーテルに溶解し、常法により 処理した。残渣をn−ヘキサンにより 結晶化し、さらに酢酸エチル・n−ヘ キサンから再結晶を行ない製品とした。Dissolve the residue in ether and use the usual method Processed. The residue was diluted with n-hexane. Crystallized and further diluted with ethyl acetate/n-hexyl The product was recrystallized from xane.
収量8.099 、融点145〜147℃0Rfl O
,? 8 、 Rfl O,82゜〔α〕腎−85,9
°(a−i、4g、メタノール)元素分析 084H,
、N30.。として計算値 061.52. H8,
05,N 6.33実験値 061..87. H7
,99,N 6.47(3)z−工1e −Val −
Asp (0But ) −Glu(onut)。Yield 8.099, melting point 145-147℃0RflO
,? 8, Rfl O, 82° [α] Kidney-85, 9
°(ai, 4g, methanol) elemental analysis 084H,
, N30. . Calculated value as 061.52. H8,
05, N 6.33 Experimental value 061. .. 87. H7
,99,N 6.47(3)z-Work1e -Val-
Asp(0But)-Glu(onut).
Z −Val−Aop (0But ) −Glu (
0But )。Z -Val-Aop (0But) -Glu (
0But).
5.349をメタノール中でパラジウム黒を触媒として
20時間接接触光した
( Rfl O,68、RfIIo、76 )。濾過し
溶媒を留去したのち残渣を減圧乾燥し
た。5.349 was catalyzed in methanol with palladium black as a catalyst for 20 hours (RflO, 68, RfIIo, 76). After filtration and distilling off the solvent, the residue was dried under reduced pressure.
Z−11e−OH2,189をT HIF50−に溶か
した溶液にN−メチルモ
ルホリン0.885−及びりpルギ酸イソブチル1.0
6−を−18℃で攪拌しながら加えたのち、これに上記
還元体
のN−メチルモルホリン(0,885mg)を含むTH
F(and)溶液を一18℃で攪拌しながら加えた。−
10℃で1
分間攪拌したのち、溶媒を留去し酢酸
エチルに溶解した。常法処理により得
られた酢酸エチル抽出物を石油エーテ
ルにより固化し、酢酸エチル・n−ヘ
キサンから再結晶を行ない製品とした。In a solution of Z-11e-OH2,189 dissolved in T HIF50-, 0.885-N-methylmorpholine and 1.0-isobutyl p-purformate were added.
After adding 6- with stirring at -18°C, TH containing the above reduced N-methylmorpholine (0,885 mg) was added.
The F(and) solution was added with stirring at -18°C. −
After stirring at 10°C for 1 minute, the solvent was distilled off and the mixture was dissolved in ethyl acetate. The ethyl acetate extract obtained by conventional treatment was solidified with petroleum ether and recrystallized from ethyl acetate/n-hexane to obtain a product.
収量fi、459.融点181〜182℃。Yield fi, 459. Melting point: 181-182°C.
Rfl O,8Q 、 RflI O,98。Rfl O,8Q , RflI O,98.
〔α〕腎−48,2°(0−LO8,メタノール)
元素分析 040H64N4011として計算値 06
1.84. H8,30,Bl 7.21実験値 0
61.53. H8,16,N 7.21(4)
Z −Thr −Gly−工Re −Val −Asp
(0But )−Glu(OBut)2
2−よりe −Mal −Asp (0But ) −
Glu (onut)。[α] Kidney -48.2° (0-LO8, methanol) Elemental analysis Calculated value as 040H64N4011 06
1.84. H8,30,Bl 7.21 Experimental value 0
61.53. H8, 16, N 7.21 (4)
Z -Thr -Gly-TechRe -Val -Asp
(0But)-Glu(OBut)2 2-from e-Mal-Asp (0But)-
Glu (onut).
2.529をメタノール中パラジウム黒を触媒として2
4時間接触還元した。2.529 in methanol with palladium black as catalyst 2
Contact reduction was carried out for 4 hours.
濾過し、溶媒を留去したのち残渣をn
−ヘキサンで処理し、減圧乾燥して固
化物を得た( Rfl O,64、Rfn O,82)
Z−Thr−Gly−NHNH21,269をDMF
15−に溶かした溶液に6N−塩酸/ジオキサン1.9
4−及び亜硝酸イソアミル0.52@tを一18℃で攪
拌しながら加えた。トリエチルアミンで。After filtering and distilling off the solvent, the residue was treated with n-hexane and dried under reduced pressure to obtain a solidified product (Rfl O, 64, Rfn O, 82).
Z-Thr-Gly-NHNH21,269 in DMF
1.9 of 6N hydrochloric acid/dioxane in a solution dissolved in 15-
4- and 0.52@t of isoamyl nitrite were added at -18 DEG C. with stirring. with triethylamine.
中和したのち、上記還元体のトリエチ
ルアミン(o、45−)を含むDMF
(2o−)溶液を、加え、−15℃で2時間、さらに4
℃で18時間攪拌した。After neutralization, a DMF (2o-) solution containing the above reduced triethylamine (o, 45-) was added, and the mixture was heated at -15°C for 2 hours and further for 4 hours.
Stirred at ℃ for 18 hours.
溶媒を留去したのち、残渣にIN−ク エン酸を加えて沈澱物を得、メタノー ル・エーテルから再結晶を行ない製品 とした。収i2.559.融点216〜218℃。After distilling off the solvent, the residue was Add enoic acid to obtain a precipitate, and add methanol Products obtained by recrystallization from Le Ether And so. Yield i2.559. Melting point 216-218°C.
Rfl O,92、Rf画 0.94゜〔α〕曾−2
2.6°(0−1,13,DMF)元素分析 04el
H14’6014として計算値 059.0B、 I
(7,9B、 N 8.99実験値 058.83.
H7,92,N 8.96酸加水分解物中のアミノ
酸の比率
Asp 1.04 、 Thr 1.08 。Rfl O,92, Rf picture 0.94゜[α]曾-2
2.6° (0-1,13,DMF) elemental analysis 04el
Calculated value as H14'6014 059.0B, I
(7,9B, N 8.99 experimental value 058.83.
H7,92,N 8.96Ratio of amino acids in acid hydrolyzate Asp 1.04, Thr 1.08.
Glu 1.04 、 Gly 1.09 。Glu 1.04, Gly 1.09.
Val O,93、工1eo、86
例2: Z−Arg(NO,)−Arg−Ala−P
rO−GLn−Thr −Gly−工is−’Val−
Asp(OBut)−Glu(ODut )tの製法
Z −Thr −Gly−工Re −Val −Asp
(0But) −Glu(OBut)21.129を
メタノール(5o−)及びDMF (10*)の混液中
でパラジウム黒を触媒として18時間接触還元した。濾
過後溶媒を留去し、残渣を五酸化リン上で減圧乾燥して
固化物を得た( Rfl O,67、Rfllo、79
)。Val O, 93, Eng 1eo, 86 Example 2: Z-Arg(NO,)-Arg-Ala-P
rO-GLn-Thr -Gly-Engineering is-'Val-
Production method of Asp(OBut)-Glu(ODut)t Z -Thr -Gly-Re -Val -Asp
(0But) -Glu(OBut) 21.129 was catalytically reduced in a mixture of methanol (5o-) and DMF (10*) using palladium black as a catalyst for 18 hours. After filtration, the solvent was distilled off, and the residue was dried under reduced pressure over phosphorus pentoxide to obtain a solidified product (Rfl O, 67, Rfllo, 79
).
z −Arg (No2) −Arg −Ala −P
ro −Gin −NHNHBOI:!1.829をト
リフルオロ酢酸4艷に溶解し。z -Arg (No2) -Arg -Ala -P
ro -Gin -NHNHBOI:! Dissolve 1.829 in trifluoroacetic acid.
室温で45分間放置したのち、エーテルにより沈澱させ
ヒドラジドを得た( Rf’ 0.1 G 。After standing at room temperature for 45 minutes, it was precipitated with ether to obtain hydrazide (Rf' 0.1 G).
Rf’ 0.69 ) 。コf) ヒドラジドをDMF
lofi(に溶解した溶液に6N−塩酸/ジオキサン0
.72−及び亜硝酸イソアミ/L−0,198−を加え
一18℃で攪拌した。トリエチルアミンで中和したのち
、トリエチルアミン(0,ioa+y)を含むDMF
(20m)に溶解した上記還元体の溶液を一10℃で攪
拌しながら加えた。Rf'0.69). f) Hydrazide in DMF
Lofi (6N-hydrochloric acid/dioxane solution dissolved in 0
.. 72- and isoamide nitrite/L-0,198- were added and stirred at -18°C. After neutralization with triethylamine, DMF containing triethylamine (0,ioa+y)
A solution of the above reductant dissolved in (20ml) was added at -10°C with stirring.
2時間攪拌したのち、さらに20時間4℃で攪拌した。After stirring for 2 hours, the mixture was further stirred at 4°C for 20 hours.
溶媒を留去し、残渣を水により固化させ粗生成物を得た
( 1.759 )。DMF−メタノール−酢酸エチル
より再結晶を行ない製品とした。収J1.23.融点2
02〜204℃。The solvent was distilled off and the residue was solidified with water to obtain a crude product (1.759). The product was recrystallized from DMF-methanol-ethyl acetate. Collection J1.23. Melting point 2
02-204℃.
Rfl O,40、Rf” 0.’82゜〔α)26−
30.2°(0−1,02,DMF )。Rfl O, 40, Rf"0.'82゜[α)26-
30.2° (0-1,02, DMF).
元素分析 0.、H,、、N、902. ・0H300
0H・5H20として
計算値 C5θ42. H7,59,N 15.30
実験値 050.59. H6,99,N 15.5
0酸加水分解物中のアミノ酸の比率
Arg 1.87 、 Asp 1.05. ’1lh
r’ 0.97゜Glu 2.10. Pro 1.0
3. G171.01゜Ala 1.02 、 Val
1.01 、工1eo、94例8 : I3oc−
Tyr−Gly−8er−8er−8er−Arg−A
rg −Ala −Pro −G171− Thr −
Gly−工1e−val−Asp(0But) −Gl
u(OBut)2の製法Z −Arg (N O2)
−Arg −Ala −Pro −Gin −Thr
−Gly−工1e −Val −Aep (OBut)
−Glu (OBut)。Elemental analysis 0. ,H,,,N,902.・0H300
Calculated value as 0H・5H20 C5θ42. H7,59,N 15.30
Experimental value 050.59. H6,99,N 15.5
0 ratio of amino acids in acid hydrolyzate Arg 1.87, Asp 1.05. '1lh
r' 0.97°Glu 2.10. Pro 1.0
3. G171.01゜Ala 1.02, Val
1.01, Eng1eo, 94 Example 8: I3oc-
Tyr-Gly-8er-8er-8er-Arg-A
rg -Ala -Pro -G171- Thr -
Gly-Eng 1e-val-Asp(0But) -Gl
Production method of u(OBut)2 Z -Arg (N O2)
-Arg -Ala -Pro -Gin -Thr
-Gly- Engineering 1e -Val -Aep (OBut)
-Glu (OBut).
0.1389をD M F (40” ) *酢酸(2
−)及び水(10m)の混液中でパラジウム黒を触媒と
して40時間接触還元した。濾過し。0.1389 to DMF (40”) *acetic acid (2
-) and water (10 m), catalytic reduction was carried out for 40 hours using palladium black as a catalyst. Filter.
溶媒を留去したのち、残渣にエーテルを加えるとガム状
物が得られ、これを凍結乾燥した( Rfl O,18
、Rfl O,65)。After evaporation of the solvent, ether was added to the residue to obtain a gum, which was lyophilized (Rfl O, 18
, Rfl O, 65).
Boc −Tyr −Gly −Ser −Ser −
Ser −NHNH70,8979をDMFIO−に溶
解した溶液に6N−塩酸/ジオキサン0.32−及び亜
硝峻イソアミル0.0864を−18℃で攪拌しながら
加えた。トリエチルアミンで中和したのち、混合物にト
リエチルアミン(0,18ml )を含むDMIF (
15m<)に溶解した上記還元体の溶液を一15℃で攪
拌しながら加えた。Boc -Tyr -Gly -Ser -Ser -
To a solution in which Ser -NHNH70,8979 was dissolved in DMFIO-, 6N-hydrochloric acid/dioxane 0.32- and nitrous isoamyl 0.0864- were added with stirring at -18°C. After neutralization with triethylamine, the mixture contained DMIF (0.18 ml) (
A solution of the above reductant dissolved in 15 m<) was added at -15° C. with stirring.
−15℃で2時間攪拌したのちさらに4℃で15時間撹
拌した。溶媒を留去したのち、残渣をエーテルより沈澱
させ、得られた沈澱物を50%酢酸水溶液に溶解し、濾
過したのち濾液を50%酢酸水溶液を用いたセファデッ
クスG−25カラムクロマトグラフイー(3φxi38
cm)により精製した。溶出ピークは薄層クロマトグラ
フィー及び280 nmにおける吸光度により求めた。After stirring at -15°C for 2 hours, the mixture was further stirred at 4°C for 15 hours. After evaporating the solvent, the residue was precipitated from ether, the resulting precipitate was dissolved in a 50% acetic acid aqueous solution, filtered, and the filtrate was subjected to Sephadex G-25 column chromatography using a 50% acetic acid aqueous solution ( 3φxi38
cm). Elution peaks were determined by thin layer chromatography and absorbance at 280 nm.
このピークフラクションを収集し凍結乾燥して製品とし
た。This peak fraction was collected and freeze-dried to produce a product.
収量630”9m融点240℃(分解)。Yield 630"9m Melting point 240°C (decomposed).
Rfl O,2B 、 Rfl O,73゜〔α〕甘せ
66.2°(0−1,01,50%酢酸水溶液)。Rfl O,2B, Rfl O, 73° [α] Sweetened 66.2° (0-1,01,50% acetic acid aqueous solution).
元素分析 0118H142N230!@・20H30
00H・5H,0として
計算値 050.19. 117.55. N 14
.f!3実験値 050.04. H7,16,N
14.56酸加水分解物中のアミノ酸の比率
Arg 1.98. Asp O,99,Thr 6.
82゜Ser 1.88. Glu 2.00. Pr
o 1.08+Gly 2.00. Ala 1
.96. Val 1.0 1゜■Is O,9
4、Thy O,95例4 : Z−Ann−II
ys(Tos)−Pro−4’hr−Gly−Tyr−
Gly −Ser −Ser −Ser −Arg、
−Arg −Ala −Pro −Gin −’Ihr
−Gly−工Re −Val −Asp −Glu−
OHの製法
Boa −’[’yr −Gly −Ser −Ser
−Ser −Arg −Arg −Ala −Pro
−Gin −Thr −Gly−工Re −val
−Asp(onut)−Gtu(oBut)25 a
9 rngをアニソール(0,2gLt) を含むト
リフルオロ酢酸(1−5mg)に溶解させ、室温で45
分間放置したのちエーテルから沈澱さぜた(Rflo、
o o 、 RtHO,a、5 )。Elemental analysis 0118H142N230! @・20H30
Calculated value as 00H・5H,0 050.19. 117.55. N14
.. f! 3 Experimental value 050.04. H7,16,N
14.56 Ratio of amino acids in acid hydrolyzate Arg 1.98. Asp O, 99, Thr 6.
82°Ser 1.88. Glu 2.00. Pr
o 1.08+Gly 2.00. Ala 1
.. 96. Val 1.0 1゜■Is O,9
4, Thy O, 95 Example 4: Z-Ann-II
ys(Tos)-Pro-4'hr-Gly-Tyr-
Gly-Ser-Ser-Ser-Arg,
-Arg -Ala -Pro -Gin -'Ihr
-Gly- Engineering Re -Val -Asp -Glu-
Production method of OH Boa -'['yr -Gly -Ser -Ser
-Ser -Arg -Arg -Ala -Pro
-Gin -Thr -Gly-Re -val
-Asp(onut)-Gtu(oBut)25 a
9 rng was dissolved in trifluoroacetic acid (1-5 mg) containing anisole (0.2 g Lt) and incubated at room temperature for 45 min.
After standing for a minute, it was precipitated from ether (Rflo,
o o, RtHO,a,5).
Z −Asn−Lys (−Tos) −Pro−Th
r−Gly−N2H34BB1ngのDMF(10mg
)溶液に6N−塩酸/ジオキサン0.266−及び亜硝
酸イソアミルの10%DMF溶液10−を−18℃で攪
拌しながら加えたのち、上記沈澱物のDM’F (1O
d)溶液を一15℃で攪拌しながら加えた。−1θ℃で
2時間攪拌したのちさらに4℃で18時間攪拌し、その
後、濃縮乾固した。得られた物質を50%酢酸水溶液を
用いたセフアゾ、クスG−25カラムクロマトグラフィ
ー(3φx14Qcm)により精製した。前述と同様な
方法でピークフラクションを収集し、凍結乾燥したのち
DMF−酢酸エチルから再結晶して目的物を得た。収量
52B+19.融点21(1)(分解)。Z -Asn-Lys (-Tos) -Pro-Th
1 ng of r-Gly-N2H34BB in DMF (10 mg
) 6N hydrochloric acid/dioxane 0.266- and 10% DMF solution of isoamyl nitrite 10- were added to the solution while stirring at -18°C.
d) The solution was added with stirring at -15°C. After stirring at -1θ°C for 2 hours, the mixture was further stirred at 4°C for 18 hours, and then concentrated to dryness. The obtained substance was purified by cefazo, Kusu G-25 column chromatography (3φ×14Qcm) using a 50% aqueous acetic acid solution. Peak fractions were collected in the same manner as described above, freeze-dried, and then recrystallized from DMF-ethyl acetate to obtain the desired product. Yield 52B+19. Melting point 21(1) (decomposed).
Rfl O,27、Rfl O,56゜〔α〕背−63
.7°(o−’1L50%酢酸水溶液)。Rfl O, 27, Rfl O, 56° [α] back -63
.. 7° (o-'1L 50% acetic acid aqueous solution).
元素分析 C10?H16!N5yOssS・20H8
Coo)i・6H20として
計算値 a 4,8.71. H6,48,N15J
5実験値 048.64. H6,38,N 16.
28酸加水分解物中のアミノ酸の比率
Lys O,95,Arg 2.01. ABp 1.
99゜Thr 1.7 :3. Ser 2.35.
Glu 2.(12゜Pro 2.05. Gly 3
.15. Ala 1.0 ?。Elemental analysis C10? H16! N5yOssS・20H8
Coo) Calculated value as i・6H20 a 4,8.71. H6, 48, N15J
5 Experimental value 048.64. H6, 38, N 16.
28 Ratio of amino acids in acid hydrolyzate Lys O, 95, Arg 2.01. ABp 1.
99°Thr 1.7:3. Ser 2.35.
Glu 2. (12゜Pro 2.05. Gly 3
.. 15. Ala 1.0? .
’Val 1.02 、 工1e O,94、Tyr
1.07例5 : H−Aen−Lys−Pro’
−Thr−Gly−Tyr−Gly−Ser−8er−
3er−Arg−Arg−Ala−Pro −Gln
−Thr −Gly−工1e −Val −Asp −
Glu −OHの製法
Z −Asn −I/ya (Ta2) −Pro −
’Ihr −Gly −Tyr −Gly −Ser
−Ser −Ser −Arg −Arg −Ala
−Pro −Gln−Thr −Gly−工1e−Va
l−Asp−Glu−OHI Q lng及びH−Th
r−Pro−OH120119を一50℃で攪拌しなが
ら液体アンモニア50−に溶解した。この溶液に少量の
ナトリウムを無水条件下−50℃で攪拌しながら加え、
溶液の色が青色を呈して10秒後に塩化アンモニウム1
gを加えアンモニアを除去した。残渣をIN−塩酸でp
H3に調節し、水で5−に希釈した。不溶物を遠心分離
により除去し、上清をIM−酢酸を用いたセファデック
スG−25カラムクロマトグラフイーにより精製しピー
クフラクションを収集し凍結乾燥して製品とした。収量
68・3m9.融点193°(分解)。'Val 1.02, Engineering 1e O,94, Tyr
1.07 Example 5: H-Aen-Lys-Pro'
-Thr-Gly-Tyr-Gly-Ser-8er-
3er-Arg-Arg-Ala-Pro-Gln
-Thr -Gly-Eng.1e -Val -Asp -
Production method of Glu -OH Z -Asn -I/ya (Ta2) -Pro -
'Ihr -Gly -Tyr -Gly -Ser
-Ser -Ser -Arg -Arg -Ala
-Pro -Gln-Thr -Gly-Engineering1e-Va
l-Asp-Glu-OHI Q lng and H-Th
r-Pro-OH120119 was dissolved in liquid ammonia 50°C with stirring at -50°C. Add a small amount of sodium to this solution under stirring at −50°C under anhydrous conditions.
Ammonium chloride 1 10 seconds after the solution turns blue
g was added to remove ammonia. The residue was purified with IN-hydrochloric acid.
Adjusted to H3 and diluted to 5- with water. Insoluble matter was removed by centrifugation, and the supernatant was purified by Sephadex G-25 column chromatography using IM-acetic acid, and peak fractions were collected and freeze-dried to obtain a product. Yield 68.3m9. Melting point 193° (decomposition).
Rf O,22,Rf O,54゜〔α〕賢−79
.3°(0−α66、IM−酢酸)。Rf O, 22, Rf O, 54゜[α]ken-79
.. 3° (0-α66, IM-acetic acid).
元素分析 0.、H,、H,O,・ioH,Coon・
、ll0j!¥H,Oとして
計算値 C48・55 7.08 15・874I
41゜■4° 6.79° 16.08実験値
043.62. H6,32,N 15.75酸加
水分解物中のアミノ酸の比率
1″yrO,90,工Re O,86、Val O,8
9。Elemental analysis 0. ,H,,H,O,・ioH,Coon・
,ll0j! Calculated value as ¥H, O C48・55 7.08 15・874I
41°■4° 6.79° 16.08 Experimental value 043.62. H6,32,N 15.75 Ratio of amino acids in acid hydrolyzate 1″yrO,90, Engineering Re O,86, Val O,8
9.
Ala 1.09. Gly 8.18. Pro 2
.02゜olu 1.88. Ser 2.78. T
hr 1.82゜Asp 2.00 、 Arg 2.
00 、 TJya 0.95例6: 放射性ヨウ素化
ガラス製試験管に蒸留水に溶解した例5で得たヘンエイ
コサペプチド5μ9(5μl )及びI M ’J ン
fil緩衝液(pu 7.4 ) 2 o’ptを加え
た。Ala 1.09. Gly 8.18. Pro 2
.. 02゜olu 1.88. Ser 2.78. T
hr 1.82°Asp 2.00, Arg 2.
00, TJya 0.95 Example 6: 5 μ9 (5 μl) of the heneicosapeptide obtained in Example 5 dissolved in distilled water in a radioiodized glass test tube and I M'J Nfil buffer (pu 7.4). Added 2 o'pt.
Na It6エ 1 moi及び酸化剤としてクロラミ
ンT20μg(水溶液、10μt)を加えてよく混和し
約30秒後、100/71(水溶液、 50111 )
のソジウムメタビサルファイトで反応を停止させた。こ
の混合液をセファデックスG−N。Add 1 moi of NaIt6 and 20 μg of chloramine T (aqueous solution, 10 μt) as an oxidizing agent, mix well, and after about 30 seconds, 100/71 (aqueous solution, 50111)
The reaction was stopped with sodium metabisulfite. This mixed solution was mixed with Sephadex G-N.
のカラムで1M酢酸を溶出液としてゲル濾過した。各7
ラクシヨンの放射活性を測定し。Gel filtration was performed on a column using 1M acetic acid as an eluent. 7 each
Measure the radioactivity of lactone.
そのピークフラクションを分取して125工標識ヘンエ
イコサペプチドを得た。約200 mOi/lngの比
放射活性が得られた。The peak fraction was fractionated to obtain 125-labeled heneicosapeptide. A specific radioactivity of approximately 200 mOi/lng was obtained.
例7: 抗原の製造
例5で得たヘンエイコサペプチド51n9及び牛血清ア
ルブミン101119を水L5−に溶解した。この溶液
に水溶性カルボジイミド塩酸塩10omgを0℃で攪拌
しながら加え、0.IN塩酸でpHを5.5〜6.0に
調製したのち0℃で2時間さらに4℃で20時間攪拌し
た。その後1反応混合物に酢酸を2滴加え1M酢酸を溶
出液とするセフTデ、クスG−10カラムクロマトグラ
フィーにより精製した。溶出ピークは薄層クロマトグラ
フィー及び280nmの吸光度より求めた。このピーク
フラクションを凍結乾燥して免疫用抗原を得た。Example 7: Preparation of antigen The heneicosapeptide 51n9 obtained in Example 5 and bovine serum albumin 101119 were dissolved in water L5-. 10 omg of water-soluble carbodiimide hydrochloride was added to this solution with stirring at 0°C. After adjusting the pH to 5.5 to 6.0 with IN hydrochloric acid, the mixture was stirred at 0°C for 2 hours and further at 4°C for 20 hours. Thereafter, two drops of acetic acid were added to the reaction mixture, and the mixture was purified by column chromatography using 1M acetic acid as an eluent. The elution peak was determined by thin layer chromatography and absorbance at 280 nm. This peak fraction was freeze-dried to obtain an antigen for immunization.
例8: 抗体の製造
例7で得た免疫用抗原11119(0,51119)を
生理食塩水に溶解し、これを1st(0,5s/)のフ
ロイントの補助液(complete 1rreuna
7s adjuvant)に懸濁し、家兎〔モルモット
〕に皮下注射した。2週間後回様な方法で抗原Q、5m
g(0,251119)を用い追加免疫した。3回乃至
4回の注射後10日後に採血し抗血清を得た。Example 8: Antigen 11119 (0,51119) for immunization obtained in antibody production example 7 was dissolved in physiological saline, and this was added to 1st (0,5s/) Freund's supplementary solution (complete 1rreuna).
7s adjuvant) and injected subcutaneously into domestic rabbits (guinea pigs). Two weeks later, antigen Q, 5m
g(0,251119) was used for boosting. Blood was collected 10 days after the third or fourth injection to obtain antiserum.
例9: ラジオイムノア、セイ
0、OIMリン醗緩衝液(p)l 7.4 、0.5%
牛血清アルブミン及び0.025Mエチレンジアミン四
酢酸ナトリウム含有)0.4−と検体または同じ緩衝液
に溶かした標準液0.1−および適当な濃度に希釈した
抗血清0.1−並びニ12″工標識ヘンエイコサペプチ
ド・0.1−を混合し4℃で48時間放置した。反応液
をデキストラン炭末法の常法に従って処理し、得られた
上清の放射能を計測した。第1図のAはこのようにして
得られた標準曲線である。またBはRindorkne
chtらの方法(J、Biol、Ohem、。Example 9: Radioimmunoa, Sei 0, OIM Phosphorus Buffer (p)l 7.4, 0.5%
(containing bovine serum albumin and 0.025 M sodium ethylenediaminetetraacetate) 0.4-, a sample or a standard solution 0.1- dissolved in the same buffer solution, and an antiserum 0.1- diluted to an appropriate concentration, and a 12-inch tube. Labeled heneicosapeptide 0.1- was mixed and left at 4°C for 48 hours.The reaction solution was treated according to the conventional dextran charcoal method, and the radioactivity of the resulting supernatant was measured. A is the standard curve obtained in this way. B is the standard curve obtained in this way.
The method of cht et al. (J. Biol. Ohem.
253:2769〜2776(1978)) に準じ
てヒト血清より得られた工GII′−■を標準物質とし
て用いて得られた標準曲線である。253:2769-2776 (1978)) using GII'-■ obtained from human serum as a standard substance.
図面は標準曲線である。 The drawing is a standard curve.
Claims (1)
ド[Claims] A heneicosapeptide having an amino acid sequence represented by the formula % formula %
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP57175187A JPS5965058A (en) | 1982-10-05 | 1982-10-05 | Heneicosapeptide |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP57175187A JPS5965058A (en) | 1982-10-05 | 1982-10-05 | Heneicosapeptide |
Publications (1)
Publication Number | Publication Date |
---|---|
JPS5965058A true JPS5965058A (en) | 1984-04-13 |
Family
ID=15991798
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP57175187A Pending JPS5965058A (en) | 1982-10-05 | 1982-10-05 | Heneicosapeptide |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS5965058A (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5093317A (en) * | 1989-06-05 | 1992-03-03 | Cephalon, Inc. | Treating disorders by application of insulin-like growth factor |
US5652214A (en) * | 1989-06-05 | 1997-07-29 | Cephalon, Inc. | Treating disorders by application of insulin-like growth factors and analogs |
US6693076B1 (en) | 1989-06-05 | 2004-02-17 | Cephalon, Inc. | Treating disorders by application of insulin-like growth factors and analogs |
US6723699B1 (en) | 1989-06-05 | 2004-04-20 | Cephalon, Inc. | Treating disorders by application of insulin-like growth factors and analogs |
-
1982
- 1982-10-05 JP JP57175187A patent/JPS5965058A/en active Pending
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5093317A (en) * | 1989-06-05 | 1992-03-03 | Cephalon, Inc. | Treating disorders by application of insulin-like growth factor |
US5652214A (en) * | 1989-06-05 | 1997-07-29 | Cephalon, Inc. | Treating disorders by application of insulin-like growth factors and analogs |
US5703045A (en) * | 1989-06-05 | 1997-12-30 | Cephalon, Inc. | Treating disorders by application of insulin-like growth factors and analogs |
US5776897A (en) * | 1989-06-05 | 1998-07-07 | Cephalon, Inc. | Treating disorders by application of insulin-like growth factors and analogs |
US6693076B1 (en) | 1989-06-05 | 2004-02-17 | Cephalon, Inc. | Treating disorders by application of insulin-like growth factors and analogs |
US6723699B1 (en) | 1989-06-05 | 2004-04-20 | Cephalon, Inc. | Treating disorders by application of insulin-like growth factors and analogs |
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