JP2640971B2 - Glycosphingolipid - Google Patents

Glycosphingolipid

Info

Publication number
JP2640971B2
JP2640971B2 JP26352788A JP26352788A JP2640971B2 JP 2640971 B2 JP2640971 B2 JP 2640971B2 JP 26352788 A JP26352788 A JP 26352788A JP 26352788 A JP26352788 A JP 26352788A JP 2640971 B2 JP2640971 B2 JP 2640971B2
Authority
JP
Japan
Prior art keywords
alkyl group
carbon atoms
straight
chain alkyl
glycosphingolipid
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 - Fee Related
Application number
JP26352788A
Other languages
Japanese (ja)
Other versions
JPH02111785A (en
Inventor
徹哉 古森
隆一 樋口
泰広 河野
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Daiichi Pharmaceutical Co Ltd
Original Assignee
Daiichi Pharmaceutical Co Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Daiichi Pharmaceutical Co Ltd filed Critical Daiichi Pharmaceutical Co Ltd
Priority to JP26352788A priority Critical patent/JP2640971B2/en
Publication of JPH02111785A publication Critical patent/JPH02111785A/en
Application granted granted Critical
Publication of JP2640971B2 publication Critical patent/JP2640971B2/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Landscapes

  • Saccharide Compounds (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Description

【発明の詳細な説明】 〔産業上の利用分野〕 本発明はオニヒトデ(Acanthaster planci)由来
で、大脳皮質神経細胞性生存維持作用を有するスフイン
ゴ糖脂質に関するものである。
DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to glycosphingolipids derived from Acanthaster planci and having a cerebral cortical neuronal survival maintaining effect.

〔従来の技術及びその課題〕[Conventional technology and its problems]

スフインゴ糖脂質は、動植物の細胞膜を形成する脂肪
二重層の構成成分であり、細胞の分化、相互識別などに
関与すると言われている。
Glycosphingolipids are components of a fat bilayer that forms cell membranes of animals and plants, and are said to be involved in cell differentiation, mutual recognition, and the like.

従来、棘皮動物門に属するヒトデ類からもセレブロシ
ド(セラミド−モノヘキソシド)セラミド−ジヘキソシ
ド,ガングリオシドなどのスフインゴ糖脂質が数種得ら
れているが(T.Komori etal,Libeigs Ann.Chem,1980,
653)、いずれもセラミド(長鎖塩基と脂肪酸)部の組
成を異にする混合物(分子種)のままであり、単離され
たスフインゴ糖脂質についての構造解析や生理活性の検
討はおこなわれていない。
Conventionally, several species of glycosphingolipids such as cerebroside (ceramide-monohexoside) ceramide-dihexoside and ganglioside have also been obtained from starfishes belonging to the phylum Echinoderm (T. Komori et al., Libeigs Ann. Chem, 1980,
653), both remain mixtures (molecular species) that differ in the composition of the ceramide (long-chain base and fatty acid) moiety. Structural analysis and bioactivity studies on the isolated glycosphingolipids have been conducted. Absent.

〔課題を解決するための手段〕[Means for solving the problem]

本発明者らは、海洋物産であるオニヒトデ(Acanthas
ter planci)から新規なガングリオシドを単離し、そ
の構造決定及び生理活性を検討したところ、このものは
下記式(I)で表わされ、大脳皮質神経細胞生存作用を
有するものであることを見出し、本発明を完成した。
The present inventors have reported that the sea star, Acanthas
ter planci), a novel ganglioside was isolated, and its structure was determined and its biological activity was examined. As a result, it was found that the novel ganglioside was represented by the following formula (I) and had a cerebral cortical nerve cell survival activity. The present invention has been completed.

すなわち、本発明は次の式(I) (式中、Rは炭素数10〜25、好ましくは13〜21の直鎖ア
ルキル基を、R′は炭素数10〜20、好ましくは11〜17の
直鎖アルキル基を示す) で表わされるスフインゴ糖脂質を提供するものである。
That is, the present invention provides the following formula (I) (Wherein, R represents a linear alkyl group having 10 to 25, preferably 13 to 21 carbon atoms, and R 'represents a linear alkyl group having 10 to 20, preferably 11 to 17 carbon atoms) It provides glycolipids.

本発明のスフインゴ糖脂質(I)は、例えば、オニヒ
トデをクロロホルムとアルコール類(特にメタノール)
の混合溶媒で抽出し、濃縮後溶媒分画(後述の実施例1
を参照)を行い水溶性分画を逆相及び順相カラムクロマ
トで精製することにより得ることができる。
The glycosphingolipids (I) of the present invention can be prepared, for example, by treating a starfish with chloroform and alcohols (particularly methanol)
, Extracted with a mixed solvent, concentrated and then subjected to solvent fractionation (Example 1 described later).
And the water-soluble fraction is purified by reverse-phase and normal-phase column chromatography.

本発明のスフインゴ糖脂質には、その代表的なものと
して(I)式中R=CH217CH3,R′=CH211CH3
あるAG−2−1、(I)式中R=CH219CH3、R′=
CH211CH3であるAG−2−3及び(I)式中R=CH
221CH3,R′=CH211CH3であるAG−2−5が含ま
れ、これらは逆相高速液体クロマト(HPLC)で分離可能
である。
Typical examples of the glycosphingolipids of the present invention include AG-2-1 in which R = CH 2 ) 17 CH 3 , R ′ = CH 2 ) 11 CH 3 in the formula (I); Medium R = CH 2 ) 19 CH 3 , R ′ =
AG-2-3 which is CH 2 ) 11 CH 3 and R = CH in the formula (I)
2) 21 CH 3, R ' = CH 2) contains 11 CH AG-2-5 is 3, which are separable by reverse phase high performance liquid chromatography (HPLC).

次に、上述の如くして得られた本発明のスフインゴ糖
脂質(I)について、その大脳皮質神経細胞生存維持作
用を試験した結果を示す。
Next, the results of a test on the activity of the glycosphingolipid (I) of the present invention obtained as described above to maintain the survival of cerebral cortical neurons will be described.

MTT法: ラット胎仔(16−18日目)の大脳皮質を、デイスパー
ゼとDNアーゼにより解離し、無血清培地に懸濁した。ポ
リオルニチンでコートした96穴マルチプレートに6400細
胞/ウエルの密度で播種し(90μ/ウエル)、AG−2
−5をDMSOに溶かして添加(最終濃度100μg/ml)後、
炭酸ガスインキュペーター中で20時間培養した。そして
10μのMTT溶液を添加し(最終濃度0.15mg/ml)、更
に4時間培養を続けた。100μのイソプロパノール−
0.08N HClを加え反応を停止させ、550nmを測定波長、6
90nmを参照波長とし、吸光度を測定した。この結果を下
表に示す。
MTT method: The cerebral cortex of a rat fetus (day 16-18) was dissociated with dispase and DNase and suspended in a serum-free medium. A 96-well multiplate coated with polyornithine was seeded at a density of 6400 cells / well (90 μ / well), and AG-2
-5 dissolved in DMSO and added (final concentration 100 μg / ml)
The cells were cultured in a carbon dioxide incubator for 20 hours. And
10 μM MTT * solution was added (final concentration 0.15 mg / ml) and the culture was continued for another 4 hours. 100μ of isopropanol-
0.08N HCl was added to stop the reaction.
Absorbance was measured using 90 nm as a reference wavelength. The results are shown in the table below.

この結果から明らかなように、通常の培養条件下では
時間とともに生存大脳皮質神経細胞数が減少してゆく
が、AG−2−5の添加により神経細胞が保護され、残存
する生細胞数が増えた。
As is clear from these results, the number of surviving cerebral cortical neurons decreases with time under normal culture conditions, but the addition of AG-2-5 protects the neurons and increases the number of surviving viable cells. Was.

〔発明の効果〕〔The invention's effect〕

本発明のスフインゴ糖脂質(I)は、上記の通り大脳
皮質神経細胞生存維持作用を有するので、例えば老人性
痴呆、アルツハイマー病、パーキンソン病等の神経退行
性患者の治療薬としての用途を有するものであった。
Since the glycosphingolipid (I) of the present invention has a cerebral cortical nerve cell survival maintaining effect as described above, it has use as a therapeutic drug for neurodegenerative patients such as senile dementia, Alzheimer's disease, Parkinson's disease and the like. Met.

〔実施例〕〔Example〕

次に実施例を挙げ、本発明を更に詳しく説明する。 Next, the present invention will be described in more detail with reference to examples.

実施例1. オニヒトデ52kgのクロロホルム−メタノールエキスを
水と酢酸エチル−n−ブタノール混液で分配し、水槽を
濃縮乾固後、クロロホルム−メタノール混液で抽出し
た。このクロロホルム−メタノール可溶部を逆相および
順相カラムクロマトで精製し、順相TLC上単一スポット
を示す成分(AG−2)1.2gを得た。
Example 1. 52 kg of a chloroform-methanol extract was distributed in a mixed solution of water and ethyl acetate-n-butanol, and the water tank was concentrated to dryness and extracted with a mixed solution of chloroform-methanol. The chloroform-methanol soluble portion was purified by reversed-phase and normal-phase column chromatography to obtain 1.2 g of a component (AG-2) showing a single spot on normal-phase TLC.

実施例2. 上記のようにして得られたAG−2は、IRでアミドの吸
収を示い、13C NMRではオキシ脂肪酸を有する、フイト
スフインゴシン型セラミドに特有のシグナルが見られ
た。また、13C NMRシグナル、ネガテイブ高速原子衝撃
マススペクトロメトリー(FABMS)のイオンピークか
ら、AG−2は糖5個を有するフイトスフインゴシン型ガ
ングリオシド分子種と判断され、その糖部構造は以下の
如く明らかにした。
Example 2. AG-2 obtained as described above showed amide absorption by IR, and 13 C NMR showed a signal specific to phytosphingosin-type ceramide having an oxyfatty acid. Also, from the 13 C NMR signal and the ion peak of negative fast atom bombardment mass spectrometry (FABMS), AG-2 was determined to be a phytosphingosin-type ganglioside molecular species having five sugars, and the sugar moiety structure was as follows. I made it clear.

まず、13C NMRの知見、メタノリシス、TMS化、ガスク
ロ分析の結果から構成糖はN−アセチルノイラミン酸
(NANA)、グルコース(Glc)各1モル、ガラクトース
(Gal)3モルよりなり、各糖の結合順序は、ネガテイ
ブFABMSにおけるフラグメンテーションから、ターミナ
ルヘキソース→ヘキソース→NANA→ヘキソース→ヘキソ
ースであることが判った。
First, from the findings of 13 C NMR, the results of methanolysis, TMS, and gas chromatography, the constituent sugars consisted of 1 mol each of N-acetylneuraminic acid (NANA), glucose (Glc), and 3 mol of galactose (Gal). From the fragmentation in the negative FABMS, it was found that terminal hexose → hexose → NANA → hexose → hexose.

次にAG−2の完全メチル化体から得たアルジトール−
アセテートのGC−MS、完全メチル体のmethanolyzateの
アセテートのGC−MS分析から、ターミナルヘキソフラノ
ース(1モル)、3位結合ヘキソピラノース(1モ
ル)、4位結合NANA(1モル)、4位結合ヘキソピラノ
ース(2モル)の存在が判った。
Next, alditol obtained from the fully methylated form of AG-2
From GC-MS analysis of acetate and GC-MS analysis of acetate of completely methylated methanolyzate, terminal hexofuranose (1 mol), 3-position bound hexopyranose (1 mol), 4-position bound NANA (1 mol), 4-position bound The presence of hexopyranose (2 mol) was found.

更に、AG−2をピリジン−水混液中80℃で加熱して部
分水解すると、セラミド−ラクトシド及びオリゴ糖が得
られた。このオリゴ糖はその完全メチル化体の電解脱離
マススペクトロメトリー(FDMS)、アルジトール−アセ
テートの分析の結果から、Galf(1→3)Galp(1→
4)NANAからなるトリサツカライドであった。
Further, when AG-2 was heated at 80 ° C. in a pyridine-water mixture and partially hydrolyzed, ceramide-lactoside and oligosaccharide were obtained. This oligosaccharide was found to be Galf (1 → 3) Galp (1 → 3) based on the results of electrolytic desorption mass spectrometry (FDMS) of the completely methylated product and analysis of alditol-acetate.
4) It was a trisatsucaride made of NANA.

以上の化学的知見、AG−2の13CNMRにおけるアノメリ
ック炭素のシグナルの挙動等から、AG−2は、式(I)
で表わされる化合物の混合物であると判断された。
From the above chemical findings and the behavior of the signal of the anomeric carbon in 13 CNMR of AG-2, AG-2 is represented by the formula (I)
Was determined to be a mixture of compounds represented by

AG−2: mp.153−156℃ ネガテイブFABMS(m/z)〔1537,1565,1579,1593,1607
〔M−H〕−〕13 C NMR〔δ110.5(d),104.5(d),104.2(d),10
0.8(s),96.4(d)〕 実施例3 AG−2の成分の分離はMeOH−H2O−picAを用いた逆
相HPLC〔カラム:ERC−2151,3μ溶媒:97%MeOH−picA(1
00:5)流速:1.2ml/min〕で行い、AG−2−5を単離し
た。そしてこのものの構造を13C NMR、ネガテイブFABM
S、メタノリシスにより得られる脂肪酸メチルエステル
の同定および、ネガテイブFABMSより得られる分子量等
から同定した。 picA(イオ抑制剤;商品名) また、同様にしてAG−2−1及びAG−2−3を単離し
た。
AG-2: mp.153-156 ° C. Negative FABMS (m / z) [1537,1565,1579,1593,1607
[MH]-] 13 C NMR [δ110.5 (d), 104.5 (d), 104.2 (d), 10
0.8 (s), 96.4 (d ) ] Example 3 Separation of components of AG-2 reverse phase HPLC [column using MeOH-H 2 O-picA * : ERC-2151,3μ solvent: 97% MeOH-picA (1
00: 5) Flow rate: 1.2 ml / min] to isolate AG-2-5. And the structure of this product is 13 C NMR, negative FABM
S, identification of fatty acid methyl ester obtained by methanolysis, and molecular weight obtained by negative FABMS. * PicA (Io inhibitor; trade name) AG-2-1 and AG-2-3 were similarly isolated.

Claims (3)

(57)【特許請求の範囲】(57) [Claims] 【請求項1】次の式(I) (式中、Rは炭素数10〜25の直鎖アルキル基を、R′は
炭素数10〜20の直鎖アルキル基を示す) で表わされるスフインゴ糖脂質。
1. The following formula (I) (Wherein, R represents a straight-chain alkyl group having 10 to 25 carbon atoms, and R ′ represents a straight-chain alkyl group having 10 to 20 carbon atoms).
【請求項2】Rが炭素数13〜21の直鎖アルキル基、R′
が炭素数11〜17の直鎖アルキル基である請求項第1項記
載のスフインゴ糖脂質。
2. R is a straight-chain alkyl group having 13 to 21 carbon atoms;
Is a straight-chain alkyl group having 11 to 17 carbon atoms.
【請求項3】請求項第1項記載のスフインゴ糖脂質を有
効成分とする神経退行性疾患治療薬。
[3] A therapeutic agent for neurodegenerative diseases, comprising the glycosphingolipid of claim 1 as an active ingredient.
JP26352788A 1988-10-19 1988-10-19 Glycosphingolipid Expired - Fee Related JP2640971B2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP26352788A JP2640971B2 (en) 1988-10-19 1988-10-19 Glycosphingolipid

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP26352788A JP2640971B2 (en) 1988-10-19 1988-10-19 Glycosphingolipid

Publications (2)

Publication Number Publication Date
JPH02111785A JPH02111785A (en) 1990-04-24
JP2640971B2 true JP2640971B2 (en) 1997-08-13

Family

ID=17390773

Family Applications (1)

Application Number Title Priority Date Filing Date
JP26352788A Expired - Fee Related JP2640971B2 (en) 1988-10-19 1988-10-19 Glycosphingolipid

Country Status (1)

Country Link
JP (1) JP2640971B2 (en)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005527467A (en) * 2001-08-29 2005-09-15 ネオーズ テクノロジーズ, インコーポレイテッド Novel synthetic ganglioside derivatives and compositions thereof
JP4585172B2 (en) * 2003-01-31 2010-11-24 森永乳業株式会社 Bone formation promoter
EP1603932B1 (en) * 2003-03-06 2012-10-03 Seneb Biosciences Inc. Methods and compositions for the enzymatic synthesis of gangliosides
JP2010254718A (en) * 2010-08-13 2010-11-11 Morinaga Milk Ind Co Ltd Bone formation accelerator
EP3478279A4 (en) * 2016-06-29 2020-03-04 Hadasit Medical Research Services And Development Combinations of beta-glycolipides and 4-[(2-amino-3,5-dibromophenyl)methylamino]cyclohexan-1-ol, compositions and uses thereof in the treatment of disorders associated with protein misfolding and protein aggregations

Also Published As

Publication number Publication date
JPH02111785A (en) 1990-04-24

Similar Documents

Publication Publication Date Title
DE69735851T2 (en) METHOD FOR ACTIVATING HUMAN, ANTIGEN-ACTIVATING CELLS, ACTIVATED HUMAN, ANTIGEN-ACTIVATING CELLS, AND APPLICATIONS THEREOF
DE69032206T2 (en) LIPID-A DERIVATIVES AND THEIR USE
EP0091645B1 (en) Derivatives of amides of carboxylic acids in which the n atom is substituted by a glycosyl radical, method for their preparation and their use to influence the immunogenic system
EP0609437B1 (en) Novel sphingoglycolipid and use thereof
CH663617A5 (en) PROCESS FOR THE PREPARATION OF DERIVATIVES OF INTERNAL ESTERS OF GANGLIOSIDES AND PHARMACEUTICAL COMPOSITIONS CONTAINING THESE DERIVATIVES.
CH670645A5 (en)
JP2640971B2 (en) Glycosphingolipid
DE3508356C2 (en)
DE60109279T2 (en) PHARMACEUTICAL COMPOSITIONS WITH OLIGOSACCHARIDES AND THEIR PREPARATION
WO2005116042A1 (en) Treatment and prevention of cancer with new ginsenoside derivatives
EP0300960B1 (en) Pharmaceutical compositions and lactosyl compounds, and their preparation
US4560703A (en) Clavulone derivatives, process for preparing the same, and use of said compounds
JP2640977B2 (en) Glycosphingolipids
EP2280986B1 (en) Synthetic analogues of phosphatidyl-myo-inositol mannosides with an inhibitory activity on the inflammatory response
DE69908919T2 (en) METHOD FOR PRODUCING GM3
JPH03218391A (en) Novel g-rhizoganglioside derivative
JPS604129A (en) Antitumor agent
US5625044A (en) Methods for the preparation of pure homologous series of mono to tetra fatty acyl esters of sugars and pharmaceutical formulations useful in the treatment of cancer
JPWO2007074788A1 (en) Antitumor agent and immunostimulant
EP0432309B1 (en) Sugar derivatives, process for their preparation and their use
EP0443198B1 (en) Diterpenes with immunomodulatory activity
DE2730846A1 (en) (1)-Amino-purinyl-deoxy-ribofuranuronic acid ethylamide derivs. - with circulatory, cardiac and metabolic activity
JPH09100295A (en) Novel triterpenoid saponin and therapeutic agent containing the same as active ingredient and having action of extendingneurite
DE69002383T2 (en) Composition for the healing of autoimmune diseases.
DE102020007938A1 (en) Process for the manufacture of an analgesic/antipyretic releasing substance and its use as a medicament

Legal Events

Date Code Title Description
LAPS Cancellation because of no payment of annual fees