WO1994003498A1 - Procede de preparation ameliore de glucanes solubles - Google Patents

Procede de preparation ameliore de glucanes solubles Download PDF

Info

Publication number
WO1994003498A1
WO1994003498A1 PCT/US1993/007329 US9307329W WO9403498A1 WO 1994003498 A1 WO1994003498 A1 WO 1994003498A1 US 9307329 W US9307329 W US 9307329W WO 9403498 A1 WO9403498 A1 WO 9403498A1
Authority
WO
WIPO (PCT)
Prior art keywords
glucan
soluble
particulate
phosphate
soluble phosphorylated
Prior art date
Application number
PCT/US1993/007329
Other languages
English (en)
Inventor
David L. Williams
Rose Brigid Mcnamee
Henry A. Pretus
Isaac William Browder
Original Assignee
Bioglucans, L.P.
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 Bioglucans, L.P. filed Critical Bioglucans, L.P.
Priority to AU48376/93A priority Critical patent/AU4837693A/en
Priority to EP93921176A priority patent/EP0654045A4/fr
Publication of WO1994003498A1 publication Critical patent/WO1994003498A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0009—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Glucans, e.g. polydextrose, alternan, glycogen; (alpha-1,4)(alpha-1,6)-D-Glucans; (alpha-1,3)(alpha-1,4)-D-Glucans, e.g. isolichenan or nigeran; (alpha-1,4)-D-Glucans; (alpha-1,3)-D-Glucans, e.g. pseudonigeran; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof

Definitions

  • This invention relates to an improved, highly efficient method for preparing soluble glucans.
  • the method advantageously avoids the use of a highly polar solvent such as dimethyl sulfoxide (DMSO) .
  • DMSO dimethyl sulfoxide
  • the soluble glucans prepared by the method of the invention are non-toxic and exert pronounced immunobiological responses when administered in vivo, most notably immunostimulation of macrophage activities and stimulation of hematopoietic bone marrow activity.
  • the soluble glucans also exhibit significant effects against malignant neoplasms, including melanomas and sarcomas.
  • glucan refers generically to a variety of naturally occurring homopolysaccharides or polyglucoses, including polymers such as cellulose, amylose, glycogen, laminarians, starch, etc.
  • Glucan encompasses branched and unbranched chains of glucose units linked by 1-3, 1-4, and 1-6 glucosidic bonds that may be of either the alpha or beta type.
  • particulate glucan designates a water-insoluble particulate (about 1-3 ⁇ ) polyglucose such as that derived from the cell wall of the yeast Saccharomvces cerevisiae.
  • Particulate glucan is macromolecular and comprises a closed chain of glucopyranose units united by a series of ⁇ -1-3 glucosidic linkages.
  • Particulate glucan is a potent activator of the macrophage/monocyte cell series, complement, as well as T and B cell lymphocytes. Thus, particulate glucan has profound effects on both the reticuloendothelial and immune systems. Particulate glucan has been shown to modify host resistance to a wide variety of infectious diseases (see review by DiLuzio, 1983, Trends in Pharmacol. Sci. 4 . :344-347 and references cited therein) . In addition particulate glucan has been shown to inhibit tumor growth and prolong survival in syngeneic murine tumor models (DiLuzio et al., 1979, Advances in Exp. Med. Biol. 12_1A:269-290) .
  • glucan exerts a cytostatic effect on sarcoma and melanoma cells and a proliferative effect on normal spleen and bone marrow cells (Williams et al., 1985, Hepatology 5 :198-206) .
  • particulate glucan is administered .in vivo to animals, a number of severe side effects have become apparent, the most notable being: (1) formation of granuloma (sarcoidosis) ; (2) development of hepatosplenomegaly; (3) increased susceptibility to gram-negative infections and endotoxins; (4) activation of complement (anaphylatoxin) ; (5) development of pulmonary granulomatous vasculitis; (6) development of hypotension following intravenous administration; and (7) development of microembolism when administered at high concentrations.
  • particulate glucan is administered in vivo.
  • 20% and 100% mortality were observed in mice receiving glucan at 250 and 500 mg/kg body weight respectively.
  • glucan preparation 1-3 ⁇
  • IV intravenous
  • slightly soluble neutral glucans are commercially available, these preparations are not suitable for intravenous administration because the aqueous solutions have very high viscosity and, more importantly, because their use when administered to experimental animals has inevitably been accompanied by considerable toxicity.
  • glucan phosphate soluble phosphorylated glucan
  • glucan phosphate soluble phosphorylated glucan
  • glucan phosphate soluble phosphorylated glucan
  • soluble phosphorylated glucan refers to the class of glucans solubilized by the addition of charged phosphate groups through reaction with phosphoric acid. These are the same or substantially similar to those substances as described in U.S. Patents Nos. 4,739,046; 4,761,402; 4,818,752 and 4,833,131.
  • This soluble phosphorylated glucan is non- toxic, non-i munogenic, and substantially non- pyrogenic (see U.S. Patent Nos. 4,739,046; 4,761,402; 4,818,752 and 4,833,131).
  • glucan phosphate was prepared as follows: particulate glucan or a polyglucose-protein complex was suspended in the highly polar aprotic solvent DMSO. A strong chaotropic agent, urea, was added, the mixture heated and maintained at 50-150°C with constant stirring while phosphoric acid was slowly added. Preferably, the reaction mixture was maintained at about 100°C for about 3-12 hours to increase the yield of the bioactive product. The product was isolated and the DMSO, urea, glucose, and any unreacted phosphoric acid were removed. The yield, after reaction for about 6 hours at 100°C, is stated to be about 70-90%.
  • soluble glucans in which the polygl ⁇ copyranouse chains have acquired a charged group from a non-phosphorous containing hydrolytic acid, are described in copending application Serial No. 07/649,527.
  • the soluble glucans having a charged group including such as a sulfate or nitrate group, are also capable of exerting a pronounced immunobiological effect when administered in vivo.
  • These soluble glucans immunostimulate macrophage activity with resulting activation of the immunoactive cells in the reticuloendothelial and immune systems.
  • these soluble glucans enhance hematopoietic bone marrow activity.
  • the soluble glucans were prepared as follows: particulate glucan was suspended in a solution of DMSO and urea. The concentrated mixture was heated to about 50-150°C, a concentrated hydrolytic acid, such as sulfuric or nitric acid alone or in the presence of DMSO was added and the reaction mixture was maintained at about 50-150°C with stirring. The bioactive product was isolated and the DMSO, urea, glucose and any unreacted acid were removed. After about 6 hours at 100°C, the hydrolytic acid was used alone, the yield is stated to be about 37.5%, when the hydrolytic acid was added with additional DMSO, the yield is stated to be about 98%.
  • 4,707,471 describes a water- soluble aminated /S-l-3 bound D-glucan composition and a method for preparing such composition.
  • ⁇ 1,3-D-glucan preferably curdlan or laminarian was hydrolyzed in 90% formic acid. The acid was removed by evaporation, water was added and the mixture was refluxed for an hour. The mixture was then separated on a Sephadex G- 50 column and the highest molecular weight fraction recovered and further reacted as follows. , he hydrolyzed glucan was dissolved in water containing bromine at pH 7 and allowed to stand until all the bromine was consumed (24-48 hours) . The pH was adjusted to 5.0 and the mixture dialyzed against water and freeze dried. The oxidized glucan was added to a solution of ammonium acetate or 1,6-diaminohexane in -1-
  • the hydrolyzed laminarian was dissolved in DMSO prior to acetylation and then aminated as described above.
  • WO91/03495 by Jamas describes soluble preparations of neutral glucan polymers by a method involving treatment of glucan particles with a "unique sequence of acid and alkaline treatments.”
  • Whole glucan particles were suspended in acid solution, generally about pH 1-5 at 20-100°C, preferably using an organic acid such as acetic or formic acid.
  • the acid insoluble glucan was removed and the pH adjusted to pH 7-14.
  • the slurry was resuspended in hot alkali, such as NaOH or KOH at 0.1-10 N at 4-120°C.
  • the soluble glucan was recovered and further purified.
  • U.S. Patent No. 3,883,505 describes a method for improving solubility of poorly soluble or water- insoluble polysaccharides such as pachyman, explain, lentinan, etc. , using strong, hot aqueous solutions of urea, thiourea, guanidine and their N-lower alkyl derivatives.
  • U.S. Patent Nos. 3,987,166 and 3,943,247 describe, respectively, treatment of animal tumors and prevention and treatment of bacterial infections.
  • the glucans of these patents are highly viscous and difficult to prepare in aqueous solution of higher concentrations than 0.5% aqueous solution.
  • the present invention provides an improved, highly efficient method for preparing aqueous soluble glucans.
  • the method avoids the use of DMSO.
  • the method of the invention for preparing a soluble glucan comprises the steps of:
  • FIG. 1 (A-B) illustrates helical coil transition analyses.
  • Dextran (70kD) ( ⁇ - ⁇ ) served as the linear control.
  • Congo Red in sodium hydroxide (o- o) served as the negative control.
  • FIG. 1A is the helical coil transition analysis for soluble phosphorylated glucan prepared according to the method of the present invention.
  • FIG. IB is the helical coil transition analysis for soluble, phosphorylated ylucan prepared according to the prior art method.
  • FIG. 2 (A-B) illustrates 13 C-NMR spectra of soluble phosphorylated glucan.
  • FIG. 2A shows the NMR spectrum of soluble phosphorylated glucan prepared according to the prior art method.
  • FIG. 2B shows " the NMR spectrum of soluble phosphorylated glucan prepared according to the method of the present invention.
  • aqueous soluble glucan is prepared from a neutral polyglucose or polyglucose-protein complex obtained from a variety of microbial sources as follows: a neutral polyglucose or a polyglucose- protein complex is mixed with a strong chaotropic agent, such as urea, and the dry mixture is thoroughly mixed and ground into a fine powder. Any means of grinding to form a fine powder is suitable. For preparation of small batches, a mortar and pestle is satisfactory. In practice, about 1-4 gm of neutral polyglucan or a polyglucose-protein-complex is mixed with about 10-20 gm of urea.
  • Concentrated phosphoric acid about 5-50 ml (concentrated, about 20-43%) is added to form a slurry and the reaction mixture is heated to about 60-80°C with constant stirring. The reaction mixture is maintained at about 60-80°C for 1- 6 hours until a precipitate comprising the soluble phosphorylated glucan forms.
  • About 10 ml of distilled water is added to reform a slurry and the reaction mixture maintained at about 60-80°C with stirring. The addition of water is repeated several times, preferably about three times, to maintain a slurry and heating is continued. It is preferable to maintain the reaction mixture at about 60-80°C for about 1-2 hours. In practice, after reaction for about two hours, the yield is about 97%.
  • ammonia is released from the urea and the smell of . ammonia is most noticeable between about 1-2 hours after heating is begun.
  • about 1-4 gm of particulate glucan is mixed with about 10-20 gm of urea and about 20-25 ml of concentrated phosphoric acid is used to form the slurry which is treated as above.
  • the soluble phosphorylated glucan is isolated from the reaction mixture as follows: the mixture is removed from the heat and dissolved in a large volume of distilled water so that the precipitate is resuspended. The resulting solution is filtered through coarse, medium and fine sintered filters to remove any remaining precipitate. The solution is then molecularly sieved to remove all components of less than 10,000 MW. Accordingly, any urea, glucose and unreacted phosphoric acid are removed from the solution. Any suitable method known in the art for molecular sieving can be used. For example, the solution can be sieved using Spectrapor membrane dialysis tubing dialyzed against running distilled water. In another example, the solution can be sieved using a Millipore dialyzer/concentrator with a 10,000 dalton MW membrane filter and a larger volume of dialyzing solution.
  • the method of the present invention is more rapid and more efficient than the prior art method for preparing soluble glucan phosphate.
  • the time required for solubilization is reduced from 6 hours to less than two hours.
  • the new method does not require as intense heating as the prior art method.
  • the neutral polyglucose used in the present method for preparing the soluble phosphorylated glucan can be particulate glucan isolated from the cell wall of ⁇ . cerevisiae by known methods (see e.g., DiLuzio et al., 1979, Int'l J. Cancer 224:773-779: Hassid et al., 1941, J. Amer. Chem. Soc, 6J3:295-298) .
  • soluble phosphorylated glucan can be prepared from neutral polyglucose or polyglucose- protein products derived from a variety of microbial sources. A non-exhaustive list of such sources is presented in Table 1 of U.S. Patent No. 4,739,046 incorporated herein by reference.
  • aqueous soluble glucan products prepared according to the method of the present invention are non-toxic, non-pyrogenic and non-immunogenic when evaluated using the interfacial ring test.
  • the soluble phosphorylated glucan prepared using the improved method of the present invention is substantially the same in composition as that formed by the prior art method.
  • the soluble glucans prepared using the present method exert profound immunobiological responses when administered in vivo. More particularly, because they are active as biological response modifiers, the products obtained using the present method are useful for prophylaxis and therapy of infectious diseases induced by a variety of microorganisms including, but not limited to, bacteria, virus, fungi and protozoal parasites. Additionally, the soluble glucans may be used for the prevention and/or treatment of opportunistic infections in animals and man which are immunosuppressed as a result of congenital or acquired immunodeficiency.
  • the soluble glucans can be used, alone or in combination for therapy of neoplasms.
  • Routes of administration include, but are not limited to: oral, injection, including but not limited to intravenous, intraperitoneal, subcutaneous, intramuscular, and topical routes.
  • the soluble glucans can be administered in combination with water, an aqueous solution or any physiologically acceptable carrier.
  • the suspension was allowed to stand overnight and the supernatant decanted.
  • the residue was further digested twice with of 3% hydrochloric acid to a total volume of 5 1 at 100°C.
  • the residue was washed with boiling water and decanted numerous times until the residue became floceulent.
  • One 1 of ethyl alcohol was added to the residue, mixed thoroughly and allowed to stand a minimum of 24 hours for maximum extraction.
  • the dark reddish-brown alcohol supernatant was aspirated from the residue and discarded.
  • the alcohol extraction procedure was repeated until the alcohol supernatant was essentially colorless.
  • the alcohol was removed by washing the residue four times with hot water; the particulate glucan preparation was then collected by centrifugation, frozen and lyophilized.
  • Soluble phosphorylated glucan was prepared according to the present invention by solubilization and phosphorylation of the particulate glucan as follows: 18 gm of urea was mixed with 1 gm of particulate glucan and ground with a pestle in a mortar to form a finely ground powder mixture. Twenty-five ml of phosphoric acid (43%) was added slowly to the powder mixture to form a slurry. The mixture was heated to about 60-80°C and maintained at that temperature for about 1-2 hours with stirring. A precipitate was formed which became visible after about 1-1.5 hours and increased in amounts thereafter. About 10 ml of distilled water (Milli-Q water) was added to the mixture and heating continued with stirring.
  • the resulting solution containing the soluble phosphorylated glucan was then molecularly sieved to remove low molecular weight fractions, including glucose and urea.
  • the mixture was filtered through coarse (1-3 ⁇ ) , medium (0.8 ⁇ , 0.65 ⁇ ) and fine (0.45 ⁇ ) sintered Millipore filters to remove the precipitate.
  • the solution was then molecularly sieved using a Millipore dialyzer/concentrator (Millipore Corp. , Bedford, MA) with a 10,000 MW membrane filter. Dialysis against about 24-100 L of distilled water (Milli-Q grade water) was used to remove low MW compounds.
  • Glucan Phosphate-no DMSO prepared using the method of the invention as described in Section 5 above with the soluble phosphorylated glucan (designated Glucan
  • Phosphate prepared using the prior art method, i.e., the method of U.S. Patent No. 4,739,046.
  • the elemental composition of Glucan Phosphate prepared according to the method of the present invention indicates that it is essentially identical to the art Glucan Phosphate.
  • Table 1 the only difference seen between the product of the present method i.e. Glucan Phosphate-no DMSO and the art Glucan Phosphate is the degree of phosphorylation.
  • Glucan Phosphate-no DMSO has approximately one phosphate substitution for every 3 glucose units, whereas Glucan Phosphate has approximately one phosphate substitution for every 7 glucose units.
  • the molecular weight (polymer) distribution of the two glucans was determined by aqueous gel permeation chromatography (GPC) .
  • the basic GPC system consisted of a Waters 600E solvent delivery system, a U6K manual injector and a column heating chamber (Waters Chromatography Division, Millipore Corp. , Milford, MA).
  • the mobile phase 0.05 M sodium nitrite, was stored in a sterile reservoir (Kontes, Vineland NJ) , and was thoroughly degassed by sparging and blanketing with helium prior to use. Mobile phase was delivered at a flow rate of 0.5 ml/min.
  • the system was calibrated using narrow-band pullulan standards and dextran standards.
  • the glucans were dissolved in mobile phase at a concentration of 2-3 mg/ml by gentle rocking until completely hydrated (about 2-3 hrs) .
  • a 200 ⁇ l injection volume was used for all analyses.
  • Absolute molecular weights of the glucan were determined by on-line multi-angle laser light scattering (MALLS) photometry employing a Dawn-F MALLS photometer fitted with a K5 flow cell (Wyatt Technology Corp., Santa Barbara, CA) .
  • Absolute MW distribution, molecular weight moments (number-average MW, Z-average MW, weight-average MW) , peak MW, polydispersity and root mean square (rms) radius moments were established with ASTRA software (v. 2.0).
  • a differential index of refraction (dn/dc) of 0.146 cm 3 /g was assumed. Reported MWs of pullulan and dextran standards used to check column calibration showed good agreement with MALLS data.
  • Intrinsic viscosity ([ ⁇ ]) of the polymeric glucans were determined by on-line differential viscometry (d.v.). For determination of [ ⁇ ] the column eluent was passed through a Viscotek Model 200 differential refractometer/viscometer and data were analyzed with Unical software (Viscotek, Porter, TX) . Molecular weight determinations of standards using . this technique showed good agreement with MALLS data. Intrinsic viscosity of pullulan standards was determined to be in close agreement with previous data. The molecular weight averages, polydispersity, and intrinsic viscosity of glucan phosphate without DMSO are shown in Table 2. For comparison, analogous data for glucan phosphate prepared according to the method of Patent No. 4,739,046 are also presented.
  • a M n represents: number-average MW
  • M w represents: weight-average MW
  • M z represents: z-average MW
  • Phosphate and Glucan Phosphate-no DMSO indicates an ordered or triple-helical conformation. In both cases, a shift in the absorption maxima at sodium hydroxide concentration of 0.1 to about 0.4 M were observed.
  • FIELD STRENGTH 50 MH; RELAXATION DELAY: 1 second, PULSE WINDOW: 15°-20°, NUMBER OF SCANS: Glucan phosphate 694 scans
  • Glucan Phosphate-no DMSO shows peaks C4 and C5 b , which do not appear in Glucan Phosphate. It is apparent, however, that the compounds show I3 C-NMR spectra which agree well with a j ⁇ -1,3-linkage (Colson, Carbohydrate Research 71: 265 , 1979) .

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Materials Engineering (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Organic Chemistry (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention se rapporte à un procédé de préparation rapide et hautement efficace de glucanes solubles aqueuses.
PCT/US1993/007329 1992-08-05 1993-08-04 Procede de preparation ameliore de glucanes solubles WO1994003498A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU48376/93A AU4837693A (en) 1992-08-05 1993-08-04 Improved method for preparing soluble glucans
EP93921176A EP0654045A4 (fr) 1992-08-05 1993-08-04 Procede de preparation ameliore de glucanes solubles.

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US92607892A 1992-08-05 1992-08-05
US926,078 1992-08-05

Publications (1)

Publication Number Publication Date
WO1994003498A1 true WO1994003498A1 (fr) 1994-02-17

Family

ID=25452720

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1993/007329 WO1994003498A1 (fr) 1992-08-05 1993-08-04 Procede de preparation ameliore de glucanes solubles

Country Status (8)

Country Link
EP (1) EP0654045A4 (fr)
CN (1) CN1091747A (fr)
AU (1) AU4837693A (fr)
CA (1) CA2141845A1 (fr)
MX (1) MX9304728A (fr)
SI (1) SI9300415A (fr)
WO (1) WO1994003498A1 (fr)
ZA (1) ZA935649B (fr)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5488040A (en) * 1989-09-08 1996-01-30 Alpha-Beta Technology, Inc. Use of neutral soluble glucan preparations to stimulate platelet production
US5622939A (en) * 1992-08-21 1997-04-22 Alpha-Beta Technology, Inc. Glucan preparation
US5633369A (en) * 1989-09-08 1997-05-27 Alpha-Beta Technology, Inc. Method for producing soluble glucans
US5786343A (en) * 1997-03-05 1998-07-28 Immudyne, Inc. Phagocytosis activator compositions and their use
US5849720A (en) * 1989-09-08 1998-12-15 Alpha-Beta Technology, Inc. Enhancement of non-specific immune defenses by administration of underivatized, aqueous soluble glucans
US6046323A (en) * 1997-07-29 2000-04-04 The Collaborative Group, Ltd. Conformations of PPG-glucan
US6369216B1 (en) 1998-09-25 2002-04-09 Biopolymer Engineering Pharmaceutical, Inc. Very high molecular weight β-glucans
US7022685B2 (en) 1998-09-25 2006-04-04 Biopolymer Engineering, Inc. Very high molecular weight β-glucans
CN117016797A (zh) * 2023-08-14 2023-11-10 四川合泰新光生物科技有限公司 一种提高葡聚糖溶解速度的方法

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7507724B2 (en) * 2001-01-16 2009-03-24 Sloan-Kettering Institute For Cancer Research Therapy-enhancing glucan

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3883505A (en) * 1972-07-17 1975-05-13 Ajinomoto Kk Method of solubilizing polysaccharides
US4739046A (en) * 1985-08-19 1988-04-19 Luzio Nicholas R Di Soluble phosphorylated glucan
US4946450A (en) * 1989-04-18 1990-08-07 Biosource Genetics Corporation Glucan/collagen therapeutic eye shields

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3883505A (en) * 1972-07-17 1975-05-13 Ajinomoto Kk Method of solubilizing polysaccharides
US4739046A (en) * 1985-08-19 1988-04-19 Luzio Nicholas R Di Soluble phosphorylated glucan
US4946450A (en) * 1989-04-18 1990-08-07 Biosource Genetics Corporation Glucan/collagen therapeutic eye shields

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP0654045A4 *

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5811542A (en) * 1989-09-08 1998-09-22 Alpha-Beta Technology, Inc. Method for producing soluble glucans
US5532223A (en) * 1989-09-08 1996-07-02 Alpha-Beta Technology, Inc. Use of aqueous soluble glucan preparations to stimulate platelet production
US5849720A (en) * 1989-09-08 1998-12-15 Alpha-Beta Technology, Inc. Enhancement of non-specific immune defenses by administration of underivatized, aqueous soluble glucans
US5633369A (en) * 1989-09-08 1997-05-27 Alpha-Beta Technology, Inc. Method for producing soluble glucans
US5663324A (en) * 1989-09-08 1997-09-02 Alpha-Beta Technology, Inc. Method for producing underivatized, aqueous soluble β(1-3) glucan
US5488040A (en) * 1989-09-08 1996-01-30 Alpha-Beta Technology, Inc. Use of neutral soluble glucan preparations to stimulate platelet production
US5817643A (en) * 1992-08-21 1998-10-06 Alpha-Beta Technology, Inc. Underivatized, aqueous soluable β(1-3) glucan, composition and method of making same
US5783569A (en) * 1992-08-21 1998-07-21 Alpha-Beta Technology, Inc. Uses for underivatized, aqueous soluble β(1-3) glucan and compositions comprising same
US5622939A (en) * 1992-08-21 1997-04-22 Alpha-Beta Technology, Inc. Glucan preparation
US5786343A (en) * 1997-03-05 1998-07-28 Immudyne, Inc. Phagocytosis activator compositions and their use
US6046323A (en) * 1997-07-29 2000-04-04 The Collaborative Group, Ltd. Conformations of PPG-glucan
US6369216B1 (en) 1998-09-25 2002-04-09 Biopolymer Engineering Pharmaceutical, Inc. Very high molecular weight β-glucans
US7022685B2 (en) 1998-09-25 2006-04-04 Biopolymer Engineering, Inc. Very high molecular weight β-glucans
US7566704B2 (en) 1998-09-25 2009-07-28 Biopolymer Engineering, Inc. Very high molecular weight β-glucans
CN117016797A (zh) * 2023-08-14 2023-11-10 四川合泰新光生物科技有限公司 一种提高葡聚糖溶解速度的方法
CN117016797B (zh) * 2023-08-14 2024-06-04 四川合泰新光生物科技有限公司 一种提高葡聚糖溶解速度的方法

Also Published As

Publication number Publication date
ZA935649B (en) 1994-03-07
CN1091747A (zh) 1994-09-07
CA2141845A1 (fr) 1994-02-17
MX9304728A (es) 1994-05-31
EP0654045A1 (fr) 1995-05-24
AU4837693A (en) 1994-03-03
EP0654045A4 (fr) 1996-03-20
SI9300415A (en) 1994-06-30

Similar Documents

Publication Publication Date Title
US4818752A (en) Soluble phosphorylated glucan: methods and compositions for treatment of neoplastic diseases
Williams et al. Development of a water-soluble, sulfated (1→ 3)-β-D-glucan biological response modifier derived from Saccharomyces cerevisiae
Williams et al. A method for the solubilization of a (1→ 3)-β-D-glucan isolated from Saccharomyces cerevisiae
Williams et al. Development, physicochemical characterization and preclinical efficacy evaluation of a water soluble glucan sulfate derived from Saccharomyces cerevisiae
US5811542A (en) Method for producing soluble glucans
AU650626B2 (en) Method for producing soluble glucans
EP0655921B1 (fr) Nouvelle preparation de glucanes
US6046323A (en) Conformations of PPG-glucan
US4975421A (en) Soluble phosphorylated glucan: methods and compositions for wound healing
US2437518A (en) Manufacture of infusion and injection fluids
US4900722A (en) Methods and compositions for prophylactic and therapeutic treatment of infections
EP0654045A1 (fr) Procede de preparation ameliore de glucanes solubles
US5574023A (en) Intramolecularly crosslinked (1→3)-β-D-glucans
CN112533614A (zh) 多糖的乙酰化
WO1992013896A1 (fr) Glucanes solubles
EP0172559B1 (fr) Composé polysaccharide RON, sa préparation et son utilisation
JPH026761B2 (fr)
TWI763817B (zh) 用於透析的滲透活性組成物
S̆oltés et al. HPLC and 13C‐NMR study of carboxymethyl‐β‐(1→ 6)‐d‐gluco‐β‐(1→ 3)‐d‐glucan derived from saccharomyces cerevisiae
Williams et al. Application of aqueous gel permeation chromatography with in-line multi-angle laser light scattering and differential viscometry detectors for the characterization of natural product carbohydrate pharmaceuticals
CN112914105B (zh) 改善酵母可溶β-葡聚糖乳化性的方法
WO1993021193A1 (fr) Complexe de platine et agent antineoplasique
CN116284484A (zh) 一种低分子量藤茶精多糖及其提取分离方法和用途
JPH0645644B2 (ja) 化学修飾多糖及びその製造法
EP0267335A1 (fr) Dérivés de glucanes

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU BB BG BR BY CA CZ FI HU JP KR KZ LK MG MN MW NO NZ PL RO RU SD SK UA VN

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 2141845

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 1993921176

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1993921176

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1993921176

Country of ref document: EP