Preparation of antitumor agent from shellfish

Claim 2

A process for the preparation of an antitumor agent consisting essentially of water-soluble, macromolecular glycoprotein substances having a range of molecular weights within the limits of from 10,000 to 300,000 from shellfish which comprises recovering the liquid portion which comes from cooking of raw shellfish in a hot aqueous solvent or with vapor of such solvent and which is to be discarded as waste, concentrating the liquid portion thus recovered to a smaller volume or to dryness to yield a concentrate or dry powder and isolating the water-soluble, macromolecular glycoprotein substances from the concentrate or dry powder.

Claim 3

A process for the preparation of an antitumor agent consisting essentially of water-soluble macromolecular glycoprotein substances having a range of molecular weights within the limits of from 10,000 to 300,000 from shellfish which comprises recovering the liquid portion which comes from cooking of raw shellfish in a hot aqueous solvent or with vapor of such solvent and which is to be discarded as waste, concentrating the liquid portion thus recovered to a smaller volume or to dryness to yield a concentrate or dry powder, dissolving the concentrate or dry powder in an aqueous solvent and subjecting the resulting aqueous solution to a series of treatments comprising an ion-exchange chromatography with a basic anion-exchanger and a gel-filtration whereby to collect fractions which are adsorbed on the basic anion-exchanger and which have a range of molecular weights within the limits of from 10,000 to 300,000.

Claim 4

A process for the preparation of an antitumor agent consisting essentially of water-soluble macromolecular glycoprotein substances having a range of molecular weights within the limits of from 10,000 to 300,000 from shellfish which comprises recovering the liquid portion which comes from cooking of raw shellfish in a hot aqueous solvent or with vapor of such solvent and which is to be discarded as waste, concentrating the liquid portion thus recovered to a smaller volume or to dryness to yield a concentrate or dry powder, dissolving the concentrate or dry powder in an aqueous solvent and subjecting the resulting aqueous solution to a series of treatments comprising an ion-exchange chromatography with a basic anion-exchanger and a gel-filtration whereby to collect fractions which are adsorbed on the basic anion-exchanger and which have a range of molecular weights within the limits of from 10,000 to 300,000.

Claim 5

A process for the preparation of an antitumor agent consisting essentially of water-soluble, macromolecular glycoprotein substances having a range of molecular weights within the limits of from 10,000 to 300,000 from shellfish which comprises recovering the liquid portion which comes from cooking of raw shellfish in a hot aqueous solvent or with vapor of such solvent and which is to be discarded as waste, concentrating the liquid portion thus recovered to a smaller volume or to dryness to yield a concentrate or dry powder, dissolving the concentrate or dry powder in an aqueous solvent, subjecting the resulting aqueous solution to an ion-exchange chromatography with a basic anion-exchanger to adsorb water-soluble macromolecular glycoprotein substances on the exchanger followed by desorption of the substances and subjecting the desorbed substances to a gel-filtration to collect fractions having a range of molecular weights within the limits of from 10,000 to 300,000.

Claim 6

A process for the preparation of an antitumor agent consisting essentially of water-soluble, macromolecular glycoprotein substances having a range of molecular weights within the limits of from 10,000 to 300,000 from shellfish which comprises recovering the liquid portion which comes from cooking of raw shellfish in a hot aqueous solvent or with vapor of such solvent and which is to be discarded as waste, concentrating the liquid portion thus recovered to a smaller volume or to dryness to yield a concentrate or dry powder, dissolving the concentrate or dry powder in an aqueous solvent, subjecting the resulting aqueous solution to an ion-exchange chromatography with a basic anion-exchanger to adsorb water-soluble macromolecular glycoprotein substances on the exchanger followed by desorption of the substances and subjecting the desorbed substances to a gel-filtration to collect fractions having a range of molecular weights within the limits of from 10,000 to 300,000.

Claim 7

A process according to claim 3 wherein the aqueous solution of the concentrate or dry powder is purified from insoluble matters and inorganic salts contained therein before it is treated.

Claim 8

A process for the preparation of an antitumor agent consisting essentially of water-soluble macromolecular glycoprotein substances having a range of molecular weights within the limits of from 10,000 to 300,000 from shellfish which comprises recovering the liquid portion which comes from cooking of raw shellfish in a hot aqueous solvent or with vapor of such solvent and which is to be discarded as waste, concentrating the liquid portion thus recovered to a smaller volume or to dryness to yield a concentrate or dry powder, dissolving the concentrate or dry powder in an aqueous solvent, subjecting the resulting aqueous solution to a gel-filtration to collect fractions having a range of molecular weights within the limits of from 10,000 to 300,000 and subjecting the fractions thus collected to an ion-exchange chromatography with a basic anion-exchanger to adsorb water-soluble macromolecular glycoprotein substances on the exchanger followed by desorption of the substances.

Claim 9

A process for the preparation of an antitumor agent consisting essentially of water-soluble macromolecular glycoprotein substances having a range of molecular weights within the limits of from 10,000 to 300,000 from shellfish which comprises recovering the liquid portion which comes from cooking of raw shellfish in a hot aqueous solvent or with vapor of such solvent and which is to be discarded as waste, concentrating the liquid portion thus recovered to a smaller volume or to dryness to yield a concentrate or dry powder, dissolving the concentrate or dry powder in an aqueous solvent, subjecting the resulting aqueous solution to a gel-filtration to collect fractions having a range of molecular weights within the limits of from 10,000 to 300,000 and subjecting the fractions thus collected to an ion-exchange chromatography with a basic anion-exchanger to adsorb water-soluble macromolecular glycoprotein substances on the exchanger followed by desorption of the substances.

Claim 10

A process according to claim 5 wherein the aqueous solution of the concentrate or dry powder is purified from insoluble matters and inorganic salts contained therein before it is treated.

Claim 11

A process according to claim 3 or 5 wherein the desorption of the adsorbed substances on the exchanger is effected by elution with a 0.07˜0.4 M aqueous sodium chloride solution.

Claim 12

A process according to claim 3 or 5 wherein the desorption of the adsorbed substances on the exchanger is effected by elution with a 0.1˜0.38 M aqueous sodium chloride solution.

Claim 13

A process according to claim 3 or 5 wherein the basic anion-exchanger used in the ion-exchange chromatography is one having diethylaminoethyl or aminoethyl group as ionizing group.

Claim 14

An antitumor agent having no cytotoxicity but an immunopotentiating activity and consisting essentially of water-soluble, macromolecular glycoprotein substances which are recovered from the liquid portion which comes from cooking of shellfish with a hot aqueous solvent or with the vapor of such solvent and which have the following characteristics: (1) Molecular weight: About 10,000 to about 300,000 as measured by gel-filtration chromatography, (2) Ultraviolet absorption spectrum: With a characteristic absorption peak λ max H .spsb.2 O at 278 nm, (3) Infrared absorption spectrum (pelleted in KBr): With characteristic absorption peaks at 3500˜3300, 1650, 1540 and 1400 cm -1 , (4) Solubility: Soluble easily in water but insoluble in organic solvents such as methanol, ethanol and acetone, (5) Color reactions: Positive in biuret reaction, xanthoproteic reaction, phenolic reagent reaction, anthrone-sulfuric acid reaction and phenol-sulfuric acid reaction; but pseudo-positive in cysteine-sulfuric acid reaction, (6) Acidic or basic nature: Amphoteric electrolyte, (7) Substance color and appearance: White:colored solid with or without faint brown tinge, (8) Amino acids in hydrolysate: When hydrolyzed in 6 N HCl at 105°˜110° C. for 24 hours, the resulting hydrolysate contains at least the following amino acids: aspartic acid, hydroproline, threonine, serine, glutamic acid, proline, glycine, alanine, cysteine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, arginine, histidine, hydroxylysine, (9) Carbohydrates in hydrolysate: When hydrolyzed in 2 N HCl at 80°˜90° C. for 10 hours, followed by the removal of the amino acids from the hydrolysate by ion-exchange chromatography and further by hydrogenation of the amino acids-free hydrolysate, the hydrogenated hydrolysate contains at least the following carbohydrates: fructose, mannose, fucose, inositol, galactose.

Claim 15

An antitumor agent having no cytotoxicity but an immunopotentiating activity and consisting essentially of water-soluble, macromolecular glycoprotein substances which are recovered from the liquid portion which comes from cooking of shellfish with a hot aqueous solvent or with the vapor of such solvent and which have the following characteristics: (1) Molecular weight: About 10,000 to about 300,000 as measured by gel-filtration chromatography, (2) Ultraviolet absorption spectrum: With a characteristic absorption peak λ max H .spsb.2 O at 278 nm, (3) Infrared absorption spectrum (pelleted in KBr): With characteristic absorption peaks at 3500˜3300, 1650, 1540 and 1400 cm -1 , (4) Solubility: Soluble easily in water but insoluble in organic solvents such as methanol, ethanol and acetone, (5) Color reactions: Positive in biuret reaction, xanthoproteic reaction, phenolic reagent reaction, anthrone-sulfuric acid reaction and phenol-sulfuric acid reaction; but pseudo-positive in cysteine-sulfuric acid reaction, (6) Acidic or basic nature: Amphoteric electrolyte, (7) Substance color and appearance: White:colored solid with or without faint brown tinge, (8) Amino acids in hydrolysate: When hydrolyzed in 6 N HCl at 105°˜110° C. for 24 hours, the resulting hydrolysate contains at least the following amino acids: aspartic acid, hydroproline, threonine, serine, glutamic acid, proline, glycine, alanine, cysteine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, arginine, histidine, hydroxylysine, (9) Carbohydrates in hydrolysate: When hydrolyzed in 2 N HCl at 80°˜90° C. for 10 hours, followed by the removal of the amino acids from the hydrolysate by ion-exchange chromatography and further by hydrogenation of the amino acids-free hydrolysate, the hydrogenated hydrolysate contains at least the following carbohydrates: fructose, mannose, fucose, inositol, galactose.