Patent EA-003902-B1 (EA)
Title: Apoptosis inducers
Inventor: SAKAI TAKESHI
IPC: C12N9/24
Filed: 1997-01-21
Abstract: 1. An apoptosis inducer, which contains sulfated-fucose-containing polysaccharide(s) and/or degradation product(s) thereof, said sulfated-fucose-containing polysaccharide(s, having the following physicochemical properties: (a) constituting saccharide: containing uronic acid; and (b) being degraded by a fucoidanase produced by Flavobacterium sp. SA-0082 (FERM BP-5402) to thereby form at least one of the compounds selected among those represented by the following formulae (I), (II) and (III): 2. An apoptosis inducer as set forth in Claim 1 comprising sulfated-fucose-containing polysaccharide (s) and/or degradation product(s) has the following physicochemical properties: (a) constituting saccharide: substantially being free from uronic acid; and (b) substantially incapable of being degraded by a fucoidanase produced by Flavobacterium sp. SA-0082 (FERM BP-5402); (c) capable of being degraded by enzyme, produced by Alteromonas sp. SN-1009 (FERM BP-5747), making endo-degradation sulfated-fucose-containing polysaccharide; (d) forms precipitate with excess of cetylpyridine chloride at a salt concentration of nearly 1.5 M. 3. An apoptosis inducer as set forth in Claim 1 wherein said degradation product is a compound selected among those represented by the following formulae (I), (II) and (III): 4. An apoptosis inducer as set forth in Claim 1 comprising degradation product obtained by treating a sulfated-fucose-containing polysaccharide as claimed in Claim 3 with an endo-sulfated-fucose- containing polysaccharide degrading enzyme which is produced by Alteromonas sp. SN-1009 (FERM BP-5747) and acting on sulfated-fucose-containing polysaccharides degrading said sulfated-fucose-containing polysaccharides and having the following physicochemical properties: (a) constituting saccharide: substantially being free from uronic acid; and (b) substantially incapable of being degraded by a fucoidanase produced by Flavobacterium sp. SA-0082 (FERM BP-5402); said enzyme having optimum pH value: being from about pH 7 to 8; and optimum temperature: being from about 30 degree C to 35 degree C. 5. An apoptosis inducer as set forth in Claim 2 comprising degradation product which is obtained by treating a sulfated-fucose-containing polysaccharide with fucoidinaza produced by Flavobacterium sp . SA-0082 (FERM BP-5402) and is not eliminated with an ultrafiltration membrane of 100,000 in pore size. 6. An apoptosis inducer as set forth in Claim 2 comprising degradation product which is obtained by incubating Flavobacterium sp. SA-0082 (FERM BP-5402) in the presence of a sulfated-fucose-containing polysaccharide and fractionating from the culture medium thus obtained and is not eliminated with an ultrafiltration membrane of 100,000 in pore size. 7. An apoptosis inducer as set forth in Claim 2 comprising degradation product obtained by degrading a sulfated-fucose-containing polysaccharide with an acid. 8. An apoptosis inducer as set forth in Claim 7 wherein said degradation product is one having been subjected to molecular weight fractionation. 9. An apoptosis inducer according to any of Claims 1, 2, 5, 6, 7, wherein said sulfated-fucose-containing polysaccharide and/or degradation products is one originating in Kjellmaniella crassifolia, Laminaria japonica, Fucus vesiculosus, Undaria pinnatifida or Stichopus japonicus. 10. A method for inducing apoptosis, which comprises using as the active ingredient sulfated-fucose-containing polysaccharide(s) and/or degradation product(s) thereof, wherein said sulfated-fucose-containing polysaccharide is one having the following physicochemical properties: (a) constituting saccharide: containing uronic acid; and (b) being degraded by a fucoidanase produced by Flavobacterium sp. SA-0082 (FERM BP-5402) to thereby form at least one of the compounds selected among those represented by the following formulae (I), (II) and (III): 11. A method for inducing apoptosis comprising the use as an active ingredient of sulfated-fucose-containing polysaccharide(s) and/ degradation product(s) thereof, wherein said sulfated-fucose-containing polysaccharide is one having the following physicochemical properties: (a) constituting saccharide: substantially being free from uronic acid; and (b) substantially incapable of being degraded by a fucoidanase produced by Flavobacterium sp. SA-0082 (FERM BP-5402). (c) capable of being degraded by enzyme, produced by Alteromonas sp. SN-1009 (FERM BP-5747), making endo-degradation sulfated-fucose-containing polysaccharide; (d) forms precipitate with excess of cetylpyridine chloride at a salt concentration of nearly 1.5 M. 12. A method for inducing apoptosis as set forth in Claim 10 wherein said degradation product is a compound selected among those represented by the following formulae (I), (II) and (III): 13. A method for inducing apoptosis as set forth in Claim 11 wherein said degradation product(s) is one obtained by treating a sulfated-fucose-containing polysaccharide with an endo sulfated-fucose-containing polysaccharide degrading enzyme which is produced by Alteromonas sp. SN-1009 (FERM BP-5797) acting on sulfated-fucose-containing polysaccharides having the following physicochemical properties and degrading said sulfated-fucose-containing polysaccharides: (a) constituting saccharide: substantially being free from uronic acid; and (b) substantially incapable of being degraded by a fucoidanase, produced by Flavobacterium sp. SA-0082 (FERM BP-5402); said enzyme having optimum pH value: being from about pH 7 to 8; and optimum temperature: being from about 30 to 35 degree C. 14. A method for inducing apoptosis as set forth in Claim 11 comprising a use as an active ingredient of degradation product obtained by treating a sulfated-fucose-containing polysaccharide with fucoidanase, produced by Flavobacterium sp. SA-0082 (FERM BP-5402) and is not eliminated with an ultrafiltration membrane of 100,000 in pore size. 15. A method for inducing apoptosis as set forth in Claim 11 comprising a use as an active ingredient of degradation product obtained by incubating Flavobacterium sp. SA-0082 (FERM BP-5402) in the presence of a sulfated-fucose-containing polysaccharide and fractionating from the culture medium thus obtained and is not eliminated with an ultrafiltration membrane of 100,000 in pore size. 16. A method for inducing apoptosis as set forth in Claim 11 comprising a use as an active ingredient of degradation product obtained by degrading a sulfated-fucose-containing polysaccharide with an acid. 17. A method for inducing apoptosis as set forth in Claim 16 wherein said degradation product is one having been subjected to molecular weight fractionation. 18. A method for inducing apoptosis according to any of Claims 10, 11, 14, 15, 16, wherein said sulfated-fucose-containing polysaccharide and/or degradation products is one originating in Kjellmaniella crassifolia, Laminaria japonica, Fucus vesiculosus, Undaria pinnatifida or Stichopus japonicus. 19. An anticancer drug containing apoptosis inducer as claimed in any of Claims 1 to 9. 20. A carcinostatic drug containing apoptosis inducer as claimed in any of Claims 1 to 9. 21. Sulfated-fucose-containing polysaccharides having the following physicochemical properties: (a) constituting saccharide: containing uronic acid; and (b) being degraded by a fucoidanase produced by Flavobacterium sp. SA-0082 (FERM BP-5402) to thereby form at least one of the compounds selected among those represented by the following formulae (I), (II) and (III): 22. A process for producing sulfated-fucose-containing polysaccharides as set forth in Claim 21 which involves the step of treating a mixture of sulfated-fucose-containing polysaccharides originating from marine algae and holothurian with a chemical capable of aggregating acidic polysaccharides in the presence of salts and eliminating the precipitate. 23. A process for producing sulfated-fucose-containing polysaccharides as set forth in Claim 21, which involves the step of treating a mixture of sulfated-fucose-containing polysaccharides originating from marine algae and holothurian with an anion exchange resin in the presence of a divalent cation to thereby take up the target polysaccharides. 24. A process for producing sulfated-fucose-containing polysaccharides as set forth in Claim 21, which involves the step of eliminating the coexisting coloring matters by using a polysaccharide substance or a substance having an anion exchange group. 25. Sulfated-fucose-containing polysaccharides having the following physicochemical properties: (a) constituting saccharide: substantially being free from uronic acid; and (b) substantially incapable of being degraded by a fucoidanase produced by Flavobacterium sp. SA-0082 (FERM BP-5402). (c) capable of being degraded by enzyme, produced by Alteromonas sp. SN-1009 (FERM BP-5747), making endo-degradation sulfated-fucose-containing polysaccharide; (d) forms precipitate with excess of cetylpyridine chloride at a salt concentration of nearly 1.5 M. 26. A process for producing sulfated -fucose-containing polysaccharides as set forth in Claim 25, which involves the step of treating a mixture of sulfated-fucose-containing polysaccharides originating from marine algae and holothurian with a degrading enzyme capable of degrading sulfated-fucose-containing polysaccharides containing uronic acid or a microorganism having said enzyme to thereby take up the target sulfated-fucose-containing polysaccharides. 27. A process for producing sulfated-fucose-containing polysaccharides as set forth in Claim 25, which involves the step of treating a mixture of sulfated-fucose-containing polysaccharides originating from marine algae and holothurian with a chemical capable of aggregating acidic polysaccharides in the presence of salts to thereby precipitate the target polysaccharides. 28. A process for producing sulfated-fucose-containing polysaccharides as set forth in Claim 25, which involves the step of