Polysaccharides N9GI, their preparation and therapeutic compositions containing them
AssigneeTerumo Kabushiki Kaisha
InventorsMasaki Shimizu, Takeo Nomura
Life of this patent
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Abstract
A hot-water extract of the bark of Melia azadirachta L. is treated with a gel filtrating agent with a fractionation range molecular weight from 1.times.10.sup.3 -1.times.10.sup.5 to 1.times.10.sup.3 -1.5.times.10.sup.5. There is obtained polysaccharide N9GI from the first fraction of the three divided polysaccharide fractions. Polysaccharide N9GI further treated with a gel-filtrating agent with a fractionation range molecular weight from 1.times.10.sup.3 -2.times.10.sup.5 to 1.times.10.sup.3 -8.times.10.sup.5. There are obtained polysaccharide N9GIa and polysaccharide N9GIb respectively from the first and second fractions of the two divided polysaccharide fractions.
Claims20
Polysaccharide N9Gl obtained from the bark of Melia azadirachta L. which has the following characteristics:
- the freeze dried product of said polysaccharide N9GI is a white or pale yellowish brown powder;
- said polysaccharide N9GI;
- has an infrared absorption spectrum as shown in FIG. 1, wherein IR.nu..sub.max..sup.KBr cm..sup.-1 :
- 3400, 1630, 1030;
- has an ultraviolet absorption spectrum which shows no absorption maximum with end absorption only when measured in an aqueous solution;
- is soluble in water and insoluble in organic solvents selected from methanol, ethanol, acetone, ether, chloroform, ethyl acetate, benzene, and hexane;
- reacts with phenol sulfate and anthrone sulfate;
- and
- turns greenish blue by reaction with iodine;
- said polysaccharide N9GI is a mixture of polysaccharide N9GIa and of polysaccharide N9GIb;
- said polysaccharide N9GIa;
- is a neutral polysaccharide which contains as the main chain thereof an .alpha.-(1.fwdarw.4)-glucan connected to arabinose by an .alpha.-(1.fwdarw.6) bond and is composed of glucose and arabinose at a ratio of about 5:1;
- is soluble in water and insoluble in organic solvents selected from methanol, ethanol, acetone, ether, chloroform, ethyl acetate, benzene and hexane;
- reacts with phenol sulfate and anthrone sulfate;
- turns to greenish blue by reaction with iodine;
- when analyzed by column gel chromatography using dextran gel with a fractionation range molecular weight of 1.times.10.sup.3 -2.times.10.sup.5 gives a single peak with a molecular weight of about 94,000;
- has a specific rotation of (.alpha.).sub.D.sup.22 :
- -35.degree. (c=0.4, H.sub.2 O);
- has an infrared absorption spectrum as shown in FIG. 2, wherein IR.nu..sub.max..sup.KBr cm..sup.-1 :
- 3400, 2930, 1620;
- has an ultraviolet absorption spectrum which shows no absorption maximum with end absorption only when measured in an aqueous solution;
- and
- has a 100 MHz .sup.13 C nuclear magnetic resonance spectrum measured in deuterium oxide using TMS (tetramethylsilane) as the external standard as shown in FIG. 4;
- and
- said polysaccharide N9GIb;
- is a neutral polysaccharide which contains as the main chain thereof an .alpha.-(1.fwdarw.4)-glucan interspersed with .beta.-(1.fwdarw.3)-fucose and branched with .alpha.-(1.fwdarw.6)-arabinose and is composed of glucose, arabinose and fucose at a ratio of about 5:2:1;
- is soluble in water and insoluble in organic solvents selected from methanol, ethanol, acetone, ether, chloroform, ethyl acetate, benzene, and hexane;
- reacts with phenol sulfate and anthrone sulfate;
- turns greenish blue by reaction with iodine;
- when analyzed by column gel chromatography using dextran gel with a fractionation range molecular weight of 1.times.10.sup.3 -2.times.10.sup.5 gives a single peak with a molecular weight of about 21,000;
- has a specific rotation of (.alpha.).sub.D.sup.22 :
- -46.degree. C. (c=0.28, H.sub.2 O);
- has an infrared absorption spectrum as shown in FIG. 4, wherein IR.nu..sub.max..sup.KBr cm..sup.-1 :
- 3400, 2930, 1630;
- has an ultraviolet absorption spectrum which shows no absorption maximum with end absorption only when measured in an aqueous solution;
- has a 100 MHz .sup.13 C nuclear magnetic resonance spectrum measured in deuterium oxide using TMS (tetramethylsilane) as the external standard as shown in FIG. 5.
A process for preparing polysaccharide N9GI which comprises extracting the bark of Melia azadirachta L. with hot water, treating the hot water extract by means of precipitation with an alcohol or with a permeable membrane to thereby form a purified extract, dissolving the purified extract in water to form an aqueous solution of said purified extract, treating said aqueous solution with a molecular sieving agent having a fractionation range molecular weight from 1.times.10.sup.3 -1.times.10.sup.5 to 1.times.10.sup.3 -1.5.times.10.sup.5 to thereby obtain three divided polysaccharide fractions and isolating the first fraction of the three divided polysaccharide fractions;
- said polysaccharide N9GI having the following characteristics:
- the freeze dried product of said polysaccharide N9GI is a white or pale yellowish brown powder;
- said polysaccharide N9GI;
- has an infrared absorption spectrum as shown in FIG. 1, wherein IR.nu..sub.max..sup.KBr cm..sup.-1 :
- 3400, 1630, 1030;
- has an ultraviolet absorption spectrum which shows no absorption maximum with end absorption only when measured in an aqueous solution;
- is soluble in water and insoluble in organic solvents selected from methanol, ethanol, acetone, ether, chloroform, ethyl acetate, benzene, and hexane;
- reacts with phenol sulfate and anthrone sulfate;
- and turns greenish blue by reaction with iodine.
Polysaccharide N9GIa obtained from the bark of Melia azadirachta L. which has the following characteristics:
- the freeze dried product of said polysaccharide N9GIa is a white powder;
- said polysaccharide N9GIa;
- is a neutral polysaccharide which contains as the main chain thereof an .alpha.-(1.fwdarw.4)-glucan connected to arabinose by an .alpha.-(1.fwdarw.6) bond and is composed of glucose and arabinose at a ratio of about 5:1;
- is soluble in water and insoluble in organic solvents selected from methanol, ethanol, acetone, ether, chloroform, ethyl acetate, benzene and hexane;
- reacts with phenol sulfate and anthrone sulfate;
- turns to greenish blue by reaction with iodine;
- when analyzed by column gel chromatography using dextran gel with a fractionation range molecular weight of 1.times.10.sup.3 -2.times.10.sup.5 gives a single peak with a molecular weight of about 94,000;
- has a specific rotation of [.alpha]sub.D.sup.22 :
- -35.degree. (c=0.4, H.sub.2 O);
- has an infrared absorption spectrum as shown in FIG. 2, wherein IR.nu..sub.max..sup.KBr cm..sup.-1 :
- 3400, 2930, 1620;
- has an ultraviolet absorption spectrum which shows no absorption maximum with end absorption only when measured in an aqueous solution;
- and
- has a 100 MHz .sup.13 C nuclear magnetic resonance spectrum measured in deuterium oxide using TMS (tetramethylsilane) as the external standard as shown in FIG. 4.
A process for preparing polysaccharide N9GIa which comprises extracting the bark of Melia azadirachta L. with hot water and treating the hot water extract with a molecular sieve to thereby isolate said polysaccharide N9GIa having a molecular weight of about 94,000;
- said polysaccharide N9GIa having the following characteristics:
- the freeze dried product of said polysaccharide N9GI is a white powder;
- said polysaccharide N9GIa;
- is a neutral polysaccharide which contains as the main chain thereof an .alpha.-(1.fwdarw.4)-glucan connected to arabinose by an .alpha.-(1.fwdarw.6) bond and is composed of glucose and arabinose at a ratio of about 5:1;
- is soluble in water and insoluble in organic solvents selected from methanol, ethanol, acetone, ehter, chloroform, ethyl acetate, benzene and hexane;
- reacts with phenol sulfate and anthrone sulfate;
- turns to greenish blue by reaction with iodine;
- when analyzed by column gel chromatography using dextran gel with a fractionation range molecular weight of 1.times.10.sup.3 -2.times.10.sup.5 gives a single peak with a molecular weight of about 94,000;
- has a specific rotation of [.alpha]sub.D.sup.22 :
- -35.degree. (c=0.4, H.sub.2 O);
- has an infrared absorption spectrum as shown in FIG. 2, wherein IR.nu..sub.max..sup.KBr cm..sup.-1 :
- 3400, 2930, 1620;
- has an ultraviolet absorption spectrum which shows no absorption maximum with end absorption only when measured in an aqueous solution;
- and
- has a 100 MHz .sup.13 C nuclear magnetic resonance spectrum measured in deuterium oxide using TMS (tetramethylsilane) as the external standard as shown in FIG. 4.
Polysaccharide N9GIb obtained from the bark of Melia azadirachta L. which has the following characteristics:
- the freeze dried product of said polysaccharide N9GIb is a white powder;
- said polysaccharide N9GIb;
- is a neutral polysaccharide which contains as the main chain thereof an .alpha.-(1.fwdarw.4)-glucan interspersed with .beta.-(1.fwdarw.3)-fucose and branched with .alpha.-(1.fwdarw.6)-arabinose and is composed of glucose, arabinose and fucose at a ratio of about 5:2:1;
- is soluble in water and insoluble in organic solvents selected from methanol, ethanol, acetone, ether, chloroform, ethyl acetate, benzene, and hexane;
- reacts with phenol sulfate and anthrone sulfate;
- turns greenish blue by reaction with iodine;
- when analyzed by column gel chromatography using dextran gel with a fractionation range molecular weight of 1.times.10.sup.3 -2.times.10.sup.5 gives a single peak with a molecular weight of about 21,000;
- has a specific rotation of [.alpha]sub.D.sup.22 :
- -46.degree. C. (c=0.28, H.sub.2 O);
- has an infrared absorption spectrum as shown in FIG. 4, wherein IR.nu..sub.max..sup.KBr cm..sup.-1 :
- 3400, 2930, 1630;
- has an ultraviolet absorption spectrum which shows no absorption maximum with end absorption only when measured in an aqueous solution;
- has a 100 MHz .sup.13 C nuclear magnetic resonance spectrum measured in deuterium oxide using TMS (tetramethylsilane) as the external standard as shown in FIG. 5.
A process for preparing polysaccharide N9GIb which comprises extracting the bark of Melia azadirachta L. with hot water and treating the hot water extract with a molecular sieve to thereby isolate said polysaccharide N9GIb having a molecular weight of about 21,000;
- said polysaccharide N9GIb having the following characteristics:
- the freeze dried product of said polysaccharide N9GIb is a white powder;
- said polysaccharide N9GIb;
- is a neutral polysaccharide which contains as the main chain thereof an .alpha.-(1.fwdarw.4)-glucan interspersed with .beta.-(1.fwdarw.3)-fucose and branched with .alpha.-(1.fwdarw.6)-arabinose and is composed of glucose, arabinose and fucose at a ratio of about 5:2:1;
- is soluble in water and insoluble in organic solvents selected from methanol, ethanol, acetone, ether, chloroform, ethyl acetate, benzene, and hexane;
- reacts with phenol sulfate and anthrone sulfate;
- turns greenish blue by reaction with iodine;
- when analyzed by column gel chromatography using dextran gel with a fractionation range molecular weight of 1.times.10.sup.3 -2.times.10.sup.5 gives a single peak with a molecular weight of about 21,000;
- has a specific rotation of [.alpha]sub.D.sup.22 :
- -46.degree. C. (c=0.28, H.sub.2 O);
- has an infrared absorption spectrum as shown in FIG. 4, wherein IR.nu..sub.max..sup.KBr cm..sup.-1 :
- 3400, 2930, 1630;
- has an ultraviolet absorption spectrum which shows no absorption maximum with end absorption only when measured in an aqueous solution;
- has a 100 MHz .sup.13 C nuclear magnetic resonance spectrum measured in deuterium oxide using TMS (tetramethylsilane) as the external standard as shown in FIG. 5.
Dependencies are inferred from the claim text.