JP2000290296A - Calcitonin derivative - Google Patents
Calcitonin derivativeInfo
- Publication number
- JP2000290296A JP2000290296A JP11092522A JP9252299A JP2000290296A JP 2000290296 A JP2000290296 A JP 2000290296A JP 11092522 A JP11092522 A JP 11092522A JP 9252299 A JP9252299 A JP 9252299A JP 2000290296 A JP2000290296 A JP 2000290296A
- Authority
- JP
- Japan
- Prior art keywords
- calcitonin
- man
- sugar chain
- derivative
- glcnac
- 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.)
- Withdrawn
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、糖鎖を有するカル
シトニン誘導体に関するものである。詳しくは、３位ア
スパラギン残基の側鎖アミド基の窒素原子に（Ｍａｎ）
X（ＧｌｃＮＡｃ）2（但し、Ｘは１から５までの数字を
示す。）からなる糖鎖が結合している、カルシトニンま
たはエルカトニン誘導体に関するもので、さらに詳しく
は、配列表配列番号１で表されるウナギ由来カルシトニ
ン誘導体に関するものであり、血中カルシウム濃度低下
作用が向上した、新規なカルシトニン誘導体に関するも
のである。本発明は、医薬の分野に応用される。TECHNICAL FIELD The present invention relates to a calcitonin derivative having a sugar chain. Specifically, the nitrogen atom in the side chain amide group of the asparagine residue at position 3 is (Man)
It relates to a calcitonin or elcatonin derivative to which a sugar chain consisting of X (GlcNAc) 2 (where X represents a number from 1 to 5), and more specifically, represented by SEQ ID NO: 1 in the Sequence Listing The present invention relates to a calcitonin derivative derived from eel, and a novel calcitonin derivative having an improved blood calcium concentration lowering effect. The present invention is applied to the field of medicine.
【０００２】[0002]
【従来の技術】カルシトニンは、３２残基のアミノ酸か
ら成るペプチドホルモンで、哺乳動物のカルシウム調節
ホルモンとして機能し、骨吸収を抑制することから、ヒ
ト、ブタ、サケ、ウナギ由来のカルシトニンもしくはそ
の誘導体が骨粗鬆症等の治療薬として使用されている。
カルシトニンのような生理活性ペプチドを医薬として用
いる場合、体液中の分解酵素による分解等が起こり、血
中での安定性が悪く、これが大きな問題となる場合が多
い。2. Description of the Related Art Calcitonin is a peptide hormone consisting of 32 amino acids, which functions as a calcium-regulating hormone in mammals and suppresses bone resorption. Therefore, calcitonin derived from humans, pigs, salmon, and eels or a derivative thereof. Has been used as a therapeutic agent for osteoporosis and the like.
When a physiologically active peptide such as calcitonin is used as a medicine, decomposition by a decomposing enzyme in a body fluid or the like occurs, resulting in poor stability in blood, which often poses a serious problem.
【０００３】また、生理活性ペプチドでは標的とする部
位に到達する前に途中で吸収が起こることがあり、これ
が克服すべき重要な課題となっている。糖蛋白質や糖脂
質のような複合糖質の糖鎖は、細胞の基質認識、細胞間
の認識等に関わっており、また生体内物質の吸収分解や
安定性等に寄与している。従って、元々糖鎖を持たない
カルシトニンについても、糖鎖を付加することにより、
血中での安定性、吸収代謝の改善や生理活性の向上が期
待される。[0003] In the case of a physiologically active peptide, absorption may occur halfway before reaching a target site, which is an important problem to be overcome. The sugar chains of complex carbohydrates such as glycoproteins and glycolipids are involved in cell substrate recognition, intercellular recognition, etc., and also contribute to the absorption and decomposition and stability of biological substances. Therefore, even for calcitonin which originally has no sugar chain, by adding a sugar chain,
It is expected to improve blood stability, absorption and metabolism, and bioactivity.
【０００４】カルシトニン類については既に、糖鎖を付
加することにより、安定化や吸収代謝の改善あるいは投
与経路の変更等の効果が期待できるとして、稲津ら［特
開平１０−１４７５９６号（１９９８）］はウナギ由来
カルシトニンの１位から１０位までの部分ペプチドの３
位アスパラギン残基の側鎖アミド基の窒素原子にＮ−ア
セチルグルコサミンがグリコシル化している誘導体、稲
津ら［特開平１０−１４７５９８号（１９９８）］はウ
ナギ由来カルシトニンの３位アスパラギン残基の側鎖ア
ミド基の窒素原子にＮ−アセチルグルコサミンがグリコ
シル化している誘導体、また羽田ら［特開平１０−１４
７５９９号（１９９８）］はウナギ由来カルシトニンの
３位アスパラギン残基の側鎖アミド基の窒素原子に複合
型糖鎖、高マンノース型糖鎖あるいは混成型糖鎖が結合
している誘導体について報告している。しかしながら、
現時点では糖鎖構造と機能との関係は不明な部分が多
く、カルシトニンについても、どのような構造の糖鎖を
付加すれば、医薬としてより高い血中安定性、薬理作用
を持つようになるかについては明らかではない。As for calcitonins, it has already been reported that the addition of a sugar chain can be expected to have effects such as stabilization, improvement in absorption and metabolism, and change in administration route. Inazu et al. [JP-A-10-147596 (1998)]. Is the partial peptide from position 1 to position 10 of calcitonin from eel
A derivative in which N-acetylglucosamine is glycosylated at the nitrogen atom of the side chain amide group of the asparagine residue at the position 3 of the asparagine residue at the 3rd position of eel calcitonin derived from eel (JP-A-10-147598 (1998)). Derivatives in which N-acetylglucosamine is glycosylated at the nitrogen atom of the amide group, and Haneda et al.
No. 7599 (1998)] reports on a derivative in which a complex type sugar chain, a high mannose type sugar chain or a mixed type sugar chain is bonded to the nitrogen atom of the side chain amide group of the 3-position asparagine residue of eel calcitonin. I have. However,
At present, the relationship between sugar chain structure and function is largely unknown, and what kind of sugar chain can be added to calcitonin to achieve higher blood stability and pharmacological action as a drug? It is not clear about.
【０００５】[0005]
【発明が解決しようとする課題】本発明の目的は、糖鎖
を付加することにより、血中カルシウム濃度低下作用が
向上した新規なカルシトニン誘導体を提供することであ
る。SUMMARY OF THE INVENTION An object of the present invention is to provide a novel calcitonin derivative having an improved blood calcium concentration lowering action by adding a sugar chain.
【０００６】[0006]
【課題を解決するための手段】本発明者らは、配列表配
列番号１に示される３位アスパラギン残基の側鎖アミド
基の窒素原子に糖鎖が結合した新規カルシトニン誘導体
を合成し、それらのカルシウム濃度低下作用を測定した
ところ、カルシトニンと同等以上の活性があることを確
認し、本発明を完成するに至った。Means for Solving the Problems The present inventors have synthesized novel calcitonin derivatives in which a sugar chain is bonded to the nitrogen atom of the side chain amide group of the asparagine residue at position 3 shown in SEQ ID NO: 1 in the Sequence Listing, and When the calcium concentration lowering effect of was measured, it was confirmed that the activity was equal to or higher than that of calcitonin, and the present invention was completed.
【０００７】すなわち、本発明は、（１）３位アスパラ
ギン残基の側鎖アミド基の窒素原子に（Ｍａｎ）X（Ｇ
ｌｃＮＡｃ）2からなる糖鎖が結合しているカルシトニ
ンまたはエルカトニン誘導体（但し、ＭａｎはＤ−マン
ノース、ＧｌｃＮＡｃはＮ−アセチル−Ｄ−グルコサミ
ンを示し、Ｘは１から５までの数を示す。）、および
（２）配列表配列番号１で表される、３位アスパラギン
残基の側鎖アミド基の窒素原子に（Ｍａｎ）X（Ｇｌｃ
ＮＡｃ）2からなる糖鎖が結合しているウナギ由来カル
シトニン誘導体（但し、ＭａｎはＤ−マンノース、Ｇｌ
ｃＮＡｃはＮ−アセチル−Ｄ−グルコサミンを示し、Ｘ
は１から５までの数を示す。）、に関する。That is, the present invention relates to (1) adding (Man) x (G) to the nitrogen atom of the side chain amide group of the asparagine residue at the 3-position.
lcNAc) 2 a calcitonin or elcatonin derivative to which a sugar chain is bound (however, Man indicates D-mannose, GlcNAc indicates N-acetyl-D-glucosamine, and X indicates a number from 1 to 5), And (2) adding (Man) x (Glc) to the nitrogen atom of the side chain amide group of the 3-position asparagine residue represented by SEQ ID NO: 1 in the sequence listing
NAc) An eel-derived calcitonin derivative to which a sugar chain consisting of 2 is bound (however, Man is D-mannose, GI
cNAc represents N-acetyl-D-glucosamine;
Represents a number from 1 to 5. ), Concerning.
【０００８】以下、ウナギ由来カルシトニンの場合を例
として合成法を記載するが、ヒト、ブタ、サケ、ウシ、
ヒツジ、ラット等に由来するカルシトニンにおいても、
３位アスパラギン残基は保存されており、同様に合成さ
れる。また、エルカトニンにおいても、３位はアスパラ
ギン残基であり、この合成カルシトニン誘導体の場合も
同様に合成される。Hereinafter, the synthesis method will be described by taking the case of eel calcitonin as an example.
In calcitonin derived from sheep, rats, etc.,
The 3rd asparagine residue is conserved and is synthesized similarly. Elcatonin also has an asparagine residue at position 3, and the synthetic calcitonin derivative is synthesized in the same manner.
【０００９】配列表配列番号１に示す糖鎖を有するカル
シトニン誘導体の合成は如何なる方法によってもよい。
例えば、配列表配列番号２に示す、３位アスパラギン残
基に下記（化１）で示される（Ｍａｎ）6（ＧｌｃＮＡ
ｃ）2が結合したカルシトニン誘導体をグリコシダーゼ
で処理し、糖鎖構造を改変し、配列表配列番号１に示す
糖鎖を有するカルシトニン誘導体を合成できる。The synthesis of a calcitonin derivative having a sugar chain shown in SEQ ID NO: 1 in the Sequence Listing may be performed by any method.
For example, (Man) 6 (GlcNA) represented by the following (Chemical Formula 1) is added to the 3rd asparagine residue shown in SEQ ID NO: 2 in the sequence listing.
c) The calcitonin derivative to which 2 is bound is treated with glycosidase to modify the sugar chain structure, whereby a calcitonin derivative having a sugar chain shown in SEQ ID NO: 1 in the Sequence Listing can be synthesized.
【００１０】[0010]
【化１】 Embedded image
【００１１】（式中、ＭａｎはＤ−マンノース、Ｇｌｃ
ＮＡｃはＮ−アセチル−Ｄ−グルコサミンを示す。）配
列表配列番号２に示す高マンノース型糖鎖を有するカル
シトニン誘導体は如何なる方法によってもよいが、例え
ば羽田ら［特開平１０−１４７５９９号（１９９８）］
の報告した方法に準じて合成できる。(Wherein Man is D-mannose, Glc
NAc indicates N-acetyl-D-glucosamine. The calcitonin derivative having a high mannose type sugar chain represented by SEQ ID NO: 2 in the Sequence Listing may be obtained by any method. For example, Haneda et al. [Japanese Patent Application Laid-Open No. H10-147599 (1998)]
Can be synthesized according to the method reported by
【００１２】稲津ら［特開平１０−１４７５９８号（１
９９８）］の方法に従って合成したＧｌｃＮＡｃ残基を
もつウナギ由来カルシトニンを糖鎖受容体に、卵白アル
ブミンから分離した糖アミノ酸［（Ｍａｎ）6（Ｇｌｃ
ＮＡｃ）2Ａｓｎ］を糖供与体として、ムコール ヒエ
マリス（Mucor hiemalis）由来のエンド−Ｍ等のエンド
グリコシダーゼを用いた糖転移反応を利用して合成され
る。Inazu et al. [JP-A-10-147598 (1)
998)], an eel-derived calcitonin having a GlcNAc residue synthesized as described above is used as a sugar chain receptor, and a sugar amino acid [(Man) 6 (Glc) isolated from ovalbumin.
NAc) 2 Asn] as a sugar donor using a glycosyltransfer reaction using an endoglycosidase such as endo-M derived from Mucor hiemalis.
【００１３】この配列表配列番号２に示す高マンノース
型糖鎖を有するカルシトニン誘導体を、例えばAspergil
lus saitoi由来のα−マンノシダーゼで処理すると糖鎖
構造が（Ｍａｎ）5（ＧｌｃＮＡｃ）2に改変された糖鎖
を有するカルシトニン誘導体が調製できる。また、配列
表配列番号２に示す高マンノース型糖鎖を有するカルシ
トニン誘導体を、例えばJack bean由来のα−マンノシ
ダーゼで処理すると糖鎖構造が（Ｍａｎ）1（ＧｌｃＮ
Ａｃ）2に改変された糖鎖を有するカルシトニン誘導体
が調製できる。The calcitonin derivative having a high mannose type sugar chain shown in SEQ ID NO: 2 in the Sequence Listing is, for example,
When treated with α-mannosidase derived from lus saitoi, a calcitonin derivative having a sugar chain whose sugar chain structure has been modified to (Man) 5 (GlcNAc) 2 can be prepared. Further, when the calcitonin derivative having a high mannose type sugar chain shown in SEQ ID NO: 2 in the Sequence Listing is treated with α-mannosidase derived from Jack bean, for example, the sugar chain structure becomes (Man) 1 (GlcN
Ac) 2 calcitonin derivative having a modified sugar chain can be prepared.
【００１４】α−マンノシダーゼ反応液中の反応生成物
の分析は通常、高速液体クロマトグラフィー（ＨＰＬ
Ｃ）により行われる。例えば、Ｃ18の逆相系（ＯＤＳ）
カラムを用い、０．１％のトリフルオロ酢酸（ＴＦＡ）
を含む水−アセトニトリル系溶媒で展開し、２２０ｎｍ
の紫外末端吸収により検出される。生成した糖鎖を有す
るカルシトニン誘導体は公知の手段に従って容易にα−
マンノシダーゼ反応液中から分離精製することができ
る。例えば、高速液体クロマトグラフィー（ＨＰＬ
Ｃ）、ゲルロ過クロマトグラフィー、イオン交換樹脂ク
ロマトグラフィー等により反応生成物を分離し、更に濃
縮、脱塩、乾燥等を行えばよい。The analysis of the reaction product in the α-mannosidase reaction solution is usually performed by high performance liquid chromatography (HPL).
C). For example, C18 reverse phase system (ODS)
0.1% trifluoroacetic acid (TFA) using a column
Is developed with a water-acetonitrile solvent containing
Is detected by UV end absorption. The generated calcitonin derivative having a sugar chain can be easily converted to α-
It can be separated and purified from the mannosidase reaction solution. For example, high performance liquid chromatography (HPL
The reaction product may be separated by C), gel permeation chromatography, ion exchange resin chromatography or the like, and further concentrated, desalted, dried, and the like.
【００１５】[0015]
【発明の実施の形態】以下に、本発明を更に具体的に説
明するが、本発明はこれらに限定されるものではない。
尚、以下の実施例では、配列表配列番号２の誘導体をＭ
６−ＣＴ、配列表配列番号１のＸが５の誘導体をＭ５−
ＣＴ、Ｘが１の誘導体をＭ１−ＣＴ、ウナギ由来カルシ
トニンをＣＴと略記する。BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described more specifically, but the present invention is not limited thereto.
In the following examples, the derivative of SEQ ID NO: 2
6-CT, a derivative of SEQ ID NO: 1 where X is 5
CT, the derivative in which X is 1 is abbreviated as M1-CT, and eel calcitonin is abbreviated as CT.
【００１６】[0016]
【実施例１】Ｍ５−ＣＴの合成 羽田ら［特開平１０−１４７５９９号（１９９８）］の
報告した方法に準じて合成したＭ６−ＣＴ０．０８１ｍ
ｇを水１１μｌに溶解し、Aspergillus saitoi由来のα
−マンノシダーゼ（Oxford Glycosystems社製）溶液
（１μＵ／μｌ）を５μｌと酢酸ナトリウム緩衝液（５
００ｍＭ、ｐＨ５．０）を４μｌ添加し、室温で一晩イ
ンキュベートした。反応液に２０％アセトニトリル水溶
液を１００μｌ添加し希釈した後、反応生成物をＨＰＬ
Ｃ［ＯＤＳ（Ｃｏｓｍｏｓｉｌ ５Ｃ１８−３００、φ
４．６×１５０ｍｍ、ナカライテスク社製）、展開溶媒
０．１％ＴＦＡ／１８％〜７２％（４０分）アセトニト
リル水溶液、流速０．８ｍｌ／分］で２２０ｎｍの吸収
により分析した。生成物のＭ５−ＣＴは、保持時間２
２．４分（Ｍ６−ＣＴの保持時間は２２．３分）のピー
クとして検出され、その反応収率（重量収率）は５４％
であった。Example 1 Synthesis of M5-CT M6-CT 0.081m synthesized according to the method reported by Haneda et al. [JP-A-10-147599 (1998)].
g was dissolved in 11 μl of water, and α from Aspergillus saitoi was dissolved.
-5 μl of mannosidase (Oxford Glycosystems) solution (1 μU / μl) and sodium acetate buffer (5
(00 mM, pH 5.0), and incubated at room temperature overnight. After diluting the reaction solution by adding 100 μl of a 20% aqueous acetonitrile solution, the reaction product was subjected to HPL
C [ODS (Cosmosil 5C18-300, φ
4.6 × 150 mm, manufactured by Nacalai Tesque, Inc.), developing solvent 0.1% TFA / 18% to 72% (40 minutes) acetonitrile aqueous solution, flow rate 0.8 ml / min] and the absorbance at 220 nm. The product M5-CT has a retention time of 2
It was detected as a peak at 2.4 minutes (the retention time of M6-CT was 22.3 minutes), and the reaction yield (weight yield) was 54%.
Met.
【００１７】ＨＰＬＣで分取した生成物の一部を質量分
析した。ＭＡＬＤＩ−ＴＯＦＭＳ分析で、ｍ／ｚ＝４６
３２（［Ｍ＋Ｈ］+ ）に主ピークが認められ、Ｍ５−Ｃ
Ｔ（分子式Ｃ192Ｈ317Ｏ82Ｎ45Ｓ2、平均分子量４６３
２．０）であることが確認できた。A part of the product collected by HPLC was subjected to mass spectrometry. According to MALDI-TOFMS analysis, m / z = 46.
32 ([M + H] + ) with a major peak, M5-C
T (molecular formula C 192 H 317 O 82 N 45 S 2 , average molecular weight 463
2.0).
【００１８】[0018]
【実施例２】Ｍ１−ＣＴの合成 Ｍ６−ＣＴ０．０８１ｍｇを水１１μｌに溶解し、Jack
bean由来のα−マンノシダーゼ（Oxford Glycosystems
社製）溶液（１００ｍＵ／μｌ）を５μｌと酢酸ナトリ
ウム緩衝液（５００ｍＭ、ｐＨ５．０）を４μｌ添加
し、室温で一晩インキュベートした。反応液に２０％ア
セトニトリル水溶液を１００μｌ添加し希釈した後、反
応生成物をＨＰＬＣ［ＯＤＳ（Ｃｏｓｍｏｓｉｌ ５Ｃ
１８−３００、φ４．６×１５０ｍｍ、ナカライテスク
社製）、展開溶媒０．１％ＴＦＡ／１８％〜７２％（４
０分）アセトニトリル水溶液、流速０．８ｍｌ／分］で
２２０ｎｍの吸収により分析した。生成物のＭ１−ＣＴ
は、保持時間２３．４分（Ｍ６−ＣＴの保持時間は２
２．３分）のピークとして検出され、その反応収率（重
量収率）は３５％であった。Example 2 Synthesis of M1-CT 0.081 mg of M6-CT was dissolved in 11 μl of water,
α-mannosidase from bean (Oxford Glycosystems
(100 mU / μl) and 4 μl of sodium acetate buffer (500 mM, pH 5.0) were added, and the mixture was incubated at room temperature overnight. After diluting the reaction solution by adding 100 μl of a 20% aqueous acetonitrile solution, the reaction product was subjected to HPLC [ODS (Cosmosil 5C).
18-300, φ4.6 × 150 mm, manufactured by Nakarai Tesque), developing solvent 0.1% TFA / 18% to 72% (4
0 min) acetonitrile aqueous solution, flow rate 0.8 ml / min] at 220 nm. M1-CT of product
Is the retention time of 23.4 minutes (the retention time of M6-CT is 2
2.3 min), and the reaction yield (weight yield) was 35%.
【００１９】ＨＰＬＣで分取した生成物の一部を質量分
析した。ＭＡＬＤＩ−ＴＯＦＭＳ分析で、ｍ／ｚ＝３９
８３（［Ｍ＋Ｈ］+ ）に主ピークが認められ、Ｍ１−Ｃ
Ｔ（分子式Ｃ168Ｈ277Ｏ62Ｎ45Ｓ2、平均分子量３９８
３．５）であることが確認できた。A part of the product collected by HPLC was subjected to mass spectrometry. According to MALDI-TOFMS analysis, m / z = 39.
83 ([M + H] + ) with a major peak, M1-C
T (molecular formula C 168 H 277 O 62 N 45 S 2 , average molecular weight 398
3.5) was confirmed.
【００２０】[0020]
【実施例３】血中カルシウム濃度低下活性測定 ［特開平７−２２８６００号（１９９５）］の試験例に
記載の方法に従って、以下のように被験物質として実施
例１、２で製造した各物質およびＣＴの、活性を測定し
た。体重９０〜１１０ｇの健康なＳ．Ｄ．系雄性ラット
を用いた。ラットを４群に分け各群１０匹とし、次に示
すようにエルカトニン標準品（旭化成工業（株）社製）
及び被験物質を静脈内に０．２ｍｌ投与した。Example 3 Measurement of Blood Calcium Concentration-Lowering Activity According to the method described in the test example of JP-A-7-228600 (1995), each of the substances produced in Examples 1 and 2 as test substances as follows and The activity of CT was measured. A healthy S. weighing 90-110 g. D. Male rats were used. The rats were divided into 4 groups, each group comprising 10 animals, and an elcatonin standard product (manufactured by Asahi Kasei Kogyo Co., Ltd.) as shown below
And 0.2 ml of the test substance was intravenously administered.
【００２１】 第１群標準品高用量（３．６ｐｍｏｌ／ｍｌ） 第２群標準品低用量（１．８ｐｍｏｌ／ｍｌ） 第３群被験物質高用量（３．６ｐｍｏｌ／ｍｌ） 第４群被験物質低用量（１．８ｐｍｏｌ／ｍｌ） 投与１時間後に、各試験動物より、採血し、血清を分離
した。原子吸光度法により血清カルシウム濃度を測定し
た。Group 1 standard product high dose (3.6 pmol / ml) Group 2 standard product low dose (1.8 pmol / ml) Group 3 test substance high dose (3.6 pmol / ml) Group 4 test substance One hour after administration of the low dose (1.8 pmol / ml), blood was collected from each test animal and serum was separated. Serum calcium concentration was measured by the atomic absorption method.
【００２２】各群の血清カルシウム濃度を用いて、平行
線検定法により標準品に対する被験物質の相対力価を求
めた。その力価を、ＣＴの力価で割って活性比を求め、
下記表１にまとめた。Using the serum calcium concentration of each group, the relative titer of the test substance to the standard was determined by a parallel line assay. The activity ratio is determined by dividing the titer by the titer of CT,
The results are summarized in Table 1 below.
【００２３】[0023]
【表１】 [Table 1]
【００２４】[0024]
【発明の効果】本発明は、３位アスパラギン残基の側鎖
アミド基の窒素原子に（Ｍａｎ）X（ＧｌｃＮＡｃ）
2（但し、Ｘは１から５までの数字を示す。）からなる
糖鎖が結合している、血中カルシウム濃度低下作用が向
上した、カルシトニンまたはエルカトニン誘導体を提供
する。本発明は、医薬の分野に応用される。According to the present invention, (Man) x (GlcNAc) is added to the nitrogen atom of the side chain amide group of the asparagine residue at position 3.
2. A calcitonin or elcatonin derivative having an improved blood calcium concentration lowering action, to which a sugar chain consisting of 2 (where X represents a number from 1 to 5) is provided. The present invention is applied to the field of medicine.
【００２５】[0025]
【配列表】 <110> Asahi Chemical Industry Co.,Ltd. <120> カルシトニン誘導体 <130> X11-00099 <160> 2 <210> 1 <211> 32 <212> PRT <213> Anguilla japonica <220> <221> DISULFID <222> 1, 7 <220> <221> CARBOHYD <222> 3 <223> （Ｍａｎ）X（ＧｌｃＮＡｃ）2−からなる糖鎖。但し、ＭａｎはＤ−マン ノース、ＧｌｃＮＡｃはＮ−アセチル−Ｄ−グルコサミンを示し、Ｘは１から５ までの数を示す。 <220> <221> AMIDATION <222> 32 <400> 1 Cys Ser Asn Leu Ser Thr Cys Val Leu Gly Lys Leu Ser Gln Glu Leu 1 5 10 15 His Lys Leu Gln Thr Tyr Pro Arg Thr Asp Val Gly Ala Gly Thr Pro 20 25 30 <210> 2 <211> 32 <212> PRT <213> Anguilla japonica <220> <221> DISULFID <222> 1, 7 <220> <221> CARBOHYD <222> 3 <223> （Ｍａｎ）6（ＧｌｃＮＡｃ）2−からなる糖鎖。但し、ＭａｎはＤ−マン ノース、ＧｌｃＮＡｃはＮ−アセチル−Ｄ−グルコサミンを示す。 <220> <221> AMIDATION <222> 32 <400> 1 Cys Ser Asn Leu Ser Thr Cys Val Leu Gly Lys Leu Ser Gln Glu Leu 1 5 10 15 His Lys Leu Gln Thr Tyr Pro Arg Thr Asp Val Gly Ala Gly Thr Pro 20 25 30[Sequence List] <110> Asahi Chemical Industry Co., Ltd. <120> Calcitonin derivative <130> X11-00099 <160> 2 <210> 1 <211> 32 <212> PRT <213> Anguilla japonica <220><221> DISULFID <222> 1, 7 <220><221> CARBOHYD <222> 3 <223> A sugar chain comprising (Man) X (GlcNAc) 2- . Here, Man indicates D-mannose, GlcNAc indicates N-acetyl-D-glucosamine, and X indicates a number from 1 to 5. <220><221> AMIDATION <222> 32 <400> 1 Cys Ser Asn Leu Ser Thr Cys Val Leu Gly Lys Leu Ser Gln Glu Leu 1 5 10 15 His Lys Leu Gln Thr Tyr Pro Arg Thr Asp Val Gly Ala Gly Thr Pro 20 25 30 <210> 2 <211> 32 <212> PRT <213> Anguilla japonica <220><221> DISULFID <222> 1, 7 <220><221> CARBOHYD <222> 3 <223> (Man A) a sugar chain consisting of 6 (GlcNAc) 2- ; However, Man indicates D-mannose and GlcNAc indicates N-acetyl-D-glucosamine. <220><221> AMIDATION <222> 32 <400> 1 Cys Ser Asn Leu Ser Thr Cys Val Leu Gly Lys Leu Ser Gln Glu Leu 1 5 10 15 His Lys Leu Gln Thr Tyr Pro Arg Thr Asp Val Gly Ala Gly Thr Pro 20 25 30
───────────────────────────────────────────────────── フロントページの続き (72)発明者 戸▲澗▼ 一孔 東京都千代田区有楽町１丁目１番２号 旭 化成工業株式会社内 Ｆターム(参考） 4C084 AA07 BA34 BA44 DB31 NA03 ZA972 4H045 AA10 BA18 BA53 CA52 DA36 EA27 FA70 HA03 ────────────────────────────────────────────────── ─── Continuing from the front page (72) Inventor To ▲ kan ▼ 1-2-1 Yurakucho, Chiyoda-ku, Tokyo Asahi Kasei Kogyo Co., Ltd. F-term (reference) 4C084 AA07 BA34 BA44 DB31 NA03 ZA972 4H045 AA10 BA18 BA53 CA52 DA36 EA27 FA70 HA03
Claims (2)
窒素原子に（Ｍａｎ）X（ＧｌｃＮＡｃ）2からなる糖鎖
が結合しているカルシトニンまたはエルカトニン誘導
体。（但し、ＭａｎはＤ−マンノース、ＧｌｃＮＡｃは
Ｎ−アセチル−Ｄ−グルコサミンを示し、Ｘは１から５
までの数を示す。）1. A calcitonin or elcatonin derivative in which a sugar chain comprising (Man) X (GlcNAc) 2 is bonded to a nitrogen atom of a side chain amide group of an asparagine residue at position 3. (However, Man represents D-mannose, GlcNAc represents N-acetyl-D-glucosamine, and X represents 1 to 5)
Show the number up to. )
パラギン残基の側鎖アミド基の窒素原子に（Ｍａｎ）X
（ＧｌｃＮＡｃ）2からなる糖鎖が結合しているウナギ
由来カルシトニン誘導体。（但し、ＭａｎはＤ−マンノ
ース、ＧｌｃＮＡｃはＮ−アセチル−Ｄ−グルコサミン
を示し、Ｘは１から５までの数を示す。）2. The method according to claim 2, wherein (Man) X is attached to the nitrogen atom of the side chain amide group of the 3-position asparagine residue represented by SEQ ID NO: 1.
(GlcNAc) An eel-derived calcitonin derivative to which a sugar chain consisting of 2 is bound. (However, Man indicates D-mannose, GlcNAc indicates N-acetyl-D-glucosamine, and X indicates a number from 1 to 5.)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP11092522A JP2000290296A (en) | 1999-03-31 | 1999-03-31 | Calcitonin derivative |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP11092522A JP2000290296A (en) | 1999-03-31 | 1999-03-31 | Calcitonin derivative |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2000290296A true JP2000290296A (en) | 2000-10-17 |
Family
ID=14056682
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP11092522A Withdrawn JP2000290296A (en) | 1999-03-31 | 1999-03-31 | Calcitonin derivative |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2000290296A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102948732A (en) * | 2011-08-22 | 2013-03-06 | 重庆市生物技术研究所有限责任公司 | Healthcare food for increasing bone mineral density and improving osteoporosis |
-
1999
- 1999-03-31 JP JP11092522A patent/JP2000290296A/en not_active Withdrawn
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102948732A (en) * | 2011-08-22 | 2013-03-06 | 重庆市生物技术研究所有限责任公司 | Healthcare food for increasing bone mineral density and improving osteoporosis |
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