CS270103B1 - wall / l-> 6 / -beta-o-gluco- / 1 - ► 3 / -beta-o-glucan yeast - Google Patents

wall / l-> 6 / -beta-o-gluco- / 1 - ► 3 / -beta-o-glucan yeast Download PDF

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CS270103B1
CS270103B1 CS863944A CS394486A CS270103B1 CS 270103 B1 CS270103 B1 CS 270103B1 CS 863944 A CS863944 A CS 863944A CS 394486 A CS394486 A CS 394486A CS 270103 B1 CS270103 B1 CS 270103B1
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beta
gluco
wall
glucan
yeast
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CS863944A
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Czech (cs)
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CS394486A1 (en
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Ladislav Ing Csc Masler
Jozef Rndr Csc Sandula
Miroslav Ing Drsc Ferencik
Milan Clen Korespondent Repas
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Masler Ladislav
Sandula Jozef
Miroslav Ing Drsc Ferencik
Milan Clen Korespondent Repas
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Priority to CS863944A priority Critical patent/CS270103B1/en
Publication of CS394486A1 publication Critical patent/CS394486A1/en
Publication of CS270103B1 publication Critical patent/CS270103B1/en

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Abstract

stěnový /l«*6/-beta-D-glukó- (l-»-3/-beta-D-glukén kvasiniek a /06/20 + 5,0 až 8,8 0 /c«0,50 v DMSO/ molekulové hmotnosl 4xl05 až 2x10®, obaah dusíka o až 0,2 % hmot. stanovený podlá Dumasa/, obsah popola 0,3 až 0,6 % hmot., Ič-spektrum v KBr 892, 1045, 1080, 1160, 1205, 1252, 1315, 1375, 1640, 2922, 3434 CID“1, 13CNMR spektrum v DMSO-dfi so signálom 86,21 ppm. Ůčelom rieSenia je stěnový /l-*6/- -beta-D-gluko-/l-»3/-beta-D-glukán s biologicky účinnou Struktúrou, chemicky a fyzikálno chemicky definovaný, ktorý má použitie ako adjuvantná zložka vakcin a ako základna látka pre přípravu vo voda rozpustných stimulátorov nespecifických imunitných mechanizmov.wall /l«*6/-beta-D-gluco- (l-»-3/-beta-D-glucan of yeast and /06/20 + 5.0 to 8.8 0 /c«0.50 in DMSO/ molecular weightl 4xl05 to 2x10®, obaah nitrogen up to 0.2% by weight determined by Dumas/, ash content 0.3 to 0.6% by weight, IR spectrum in KBr 892, 1045, 1080, 1160, 1205, 1252, 1315, 1375, 1640, 2922, 3434 CID"1, 13CNMR spectrum in DMSO-dfi with a signal of 86.21 ppm. The purpose of the solution is wall /l-*6/- -beta-D-gluco-/l-»3/-beta-D-glucan with a biologically effective structure, chemically and physicochemically defined, which has use as an adjuvant component of vaccines and as a base substance for the preparation of water-soluble stimulators of non-specific immune mechanisms.

Description

Vynález ea týká stěnového /l-*6/-beta-D-gluko-/l-»3/-beta-D-glukánu kvasiniek.The invention relates to a yeast wall (1-) 6 -beta-D-gluco- [1- (3H) -beta-D-glucan.

Stěnové /l->6/-beta-D-gluko-/l-»»3/-beta-D-glukány tvoria skeletálnu mikrofibrilárnu část buňkových stien húb. Sú nerozpustné v organických rozpúétadlách, voda a v zriedených alkáliách, z farmakologického hlediska aú to látky netoxické, ktoré nešpecificky zvyšujú rezistenciu hostitele, voči r8znym chorobným procesom najma bakteriálnym, virusovým a hubovým infekciám, imunodeficientným atavom a neoplastickým procesorom. 3avla výrazný rádioprotektívny efekt a poeobia ako adjuvanty pri imunizácii a výrobě vakcín /G. Chihara, Y. Y. Maeda, 0. Hamuro, int. 3. Tise, Reac., 4, 207, 1982: N. R. Diluzio, Trends Pharm. Sel., 4, 344, 1983/.Wall [1-> 6] -beta-D-gluco- [1-> β-beta-D-glucans form the skeletal microfibrillar part of the cell walls of fungi. They are insoluble in organic solvents, water and dilute alkalis, from a pharmacological point of view and are non-toxic substances which non-specifically increase the resistance of the host to various disease processes, in particular bacterial, viral and fungal infections, immunodeficiency and neoplastic processors. 3avla significant radioprotective effect and poeobia as adjuvants in immunization and vaccine production / G. Chihara, Y. Y. Maeda, 0. Hamuro, int. 3. Tise, Reac., 4, 207, 1982: N. R. Diluzio, Trends Pharm. Sel., 4, 344, 1983 /.

Podstatou vynálezu je stěnový /l->6/-beta-D-gluko-/l->3/-beta-D-glukán kvasiniek s biologicky účinnou Struktúrou obecného vzorce beta-D-GlcpThe subject of the invention is a wall (1-> 6) -beta-D-gluco- (1-> 3) -beta-D-glucan of yeast with a biologically active structure of the general formula beta-D-Glcp

A-*3/-beta-D-Glcp/ /l-»3/-beta-D-Glcp)^7jQA- * 3 / -beta-D-Glcp / / l- »3 / -beta-D-Glcp) ^ 7jQ

220-1100 kde Glcp je glukopyrenozylová jednotka.220-1100 where Glcp is a glucopyrenosyl unit.

Stěnový /i-»6/-beta-D-gluko-/l~*3/-beta-D-glukán kvasiniek sa získá frakcionáciou buňkových stíén postupem podlá 0. 0. Bella a 0. H. Northcota, 3. chem. Soc. 1950, 1944 a jeho modifikáclami A. L.Houwink, 0. R. Kreger, Antonie van Leeuwenhoek 19, 1, 1953) S. Peat, W. 3. Whelan, T. E. Edwards, 3. Chem. Soc. 1958, 3862) 3. S. D. Bacon, W. C. Farmer, D. 3ones, 3. F. T. Taylor, Biochem. 3. 114, 557, 1969) D. 3. Manners, A. 3. Masson, 3. C. Patterson, 3. Gen. Microbiol. 80, 411, 1974) 3. Jelsma, D. R. Kreger, carbohyd. Res. 43, 200, 1975.Yeast wall (i-> 6) -beta-D-gluco- (1 * 3) -beta-D-glucan is obtained by fractionating cell shadows according to the procedure of 0. Bella and 0. H. Northcota, 3. chem. Soc. 1950, 1944 and its modifications A. L. Howink, 0. R. Kreger, Antonie van Leeuwenhoek 19, 1, 1953) S. Peat, W. 3. Whelan, T. E. Edwards, 3. Chem. Soc. 1958, 3862) 3. S. D. Bacon, W. C. Farmer, D. 3ones, 3. F. T. Taylor, Biochem. 3. 114, 557, 1969) D. 3. Manners, A. 3. Masson, 3. C. Patterson, 3. Gen. Microbiol. 80, 411, 1974) 3. Jelsma, D. R. Kreger, carbohyd. Res. 43, 200, 1975.

Příklad 1Example 1

100 g pekárenského droždla Saccharomyces cerevisiae sa suspenduje v 150 ml 6 % hmot, hydroxidu sodného a pri 60 °C sa mleša 4 h. suspenzia sa zriedi 100 ml 60 °C teplej vody a odstředí, sediment sa suspenduje v 150 ml 3 % hmot, hydroxidu sodného, mieša pri 80 až 100 °C 2 h a odstředí, sediment sa třikrát premyje vodou a extrahuje třikrát 20 ml 4 % hmot, kyselinou chlorovodíkovou alebo kyselinou fosforečnou za miešania pri teplote mieatnosti 2 h a odstředí, sediment sa premýva vodou dovtedy až supernatant je číry a reaguje neutrálně. Výtažok 4,5 až 5,4 % hmot, počítané na suéinu kvasiniek. Mol. hmotnost: 4 x 10S až 2 x 1O6. Glukán neobsahuje dusík a obsah popola je 0,5 % hmot. Infračervené spektrum: 892, 1045, 1080, 1160, 1205, 1252, 1315, 1375, 1640, 2922, 3434 cm1.100 g of baker's yeast Saccharomyces cerevisiae are suspended in 150 ml of 6% by weight sodium hydroxide and stirred at 60 ° C for 4 hours. The suspension is diluted with 100 ml of 60 ° C warm water and centrifuged, the sediment is suspended in 150 ml of 3% by weight. sodium hydroxide, stirred at 80-100 ° C for 2 h and centrifuged, the sediment was washed three times with water and extracted three times with 20 ml of 4% w / w, hydrochloric acid or phosphoric acid with stirring at room temperature for 2 h and centrifuged, the sediment was washed with water until the supernatant was clear and reacts neutrally. Yield 4.5 to 5.4% by weight, based on the yeast dry matter. Moth. weight: 4 x 10 S to 2 x 10 6 . Glucan does not contain nitrogen and the ash content is 0.5% by weight. Infrared Spectrum: 892, 1045, 1080, 1160, 1205, 1252, 1315, 1375, 1640, 2922, 3434 cm -1 .

Modifikovaný spdsob příkladu 1 s použitím org* kyselin a enzýmov sa uskutočni podlá známého’postupu D. 3. Manners, A. 3. Masson, 3. C. Patterson, Biochem. 3. 135, 19, 1973. .The modified method of Example 1 using org * acids and enzymes was performed according to the known procedure D. 3. Manners, A. 3. Masson, 3. C. Patterson, Biochem. 3. 135, 19, 1973.

příklad 2 ' .Example 2 '.

Perítoneálne makrofágy kontrolných morčiat /namieato stenového/l->6/-beta-D-gluko-/l-*3/-beta-D-glukánu dostali fyziologický roztok/ malí stupeň metabolickej aktivity 8,1 femtomSlov redukovaného formazánu, fagocitárnu aktivitu 64,8 % a za hodinu bolí schopné usmrtit 35,9 % přítomných patogénnych kandíd. Naproti tomu perítoneálne makrofágy morčiat, ktoré dostali 3 dávky stěnového /l->6/-beta-D-gluko-/l-*>3/-beta-D-glukánu v dňoch 12, 7, 3 dni před odberom makrofágov v množatve 10 mg/kg žívej váhy,Peritoneal macrophages of control guinea pigs (wall> /> 6) -beta-D-gluco- (1- * 3) -beta-D-glucan received saline / low metabolic activity 8.1 femtomSlov reduced formazan, phagocytic activity 64 .8% and per hour were able to kill 35.9% of the pathogenic candidates present. In contrast, peritoneal macrophages of guinea pigs that received 3 doses of wall / l-> 6 / -beta-D-gluco- / 1 - *> 3 / -beta-D-glucan on days 12, 7, 3 days before collection of macrophages in the amount 10 mg / kg body weight,

CS 270103 Bl tieto makrofágy mail stupeň metabolickej aktivity stimulovaný na 13,5 fentomólu redukovaného formazánu, rovnako bola zvýšená fagocitárna aktivita priemerne na 72,9 % a schopnost usmrcovat patogénne kandidy na 58,2CS 270103 B1 these macrophages mail the degree of metabolic activity stimulated to 13.5 phentomole of reduced formazan, as well as increased phagocytic activity increased on average to 72.9% and the ability to kill pathogenic candidiasis to 58.2

Claims (1)

PREDMET VYNÁLEZUOBJECT OF THE INVENTION Stěnový /l-^6/-beta-D-gluko-/l->3/-beta-D-glukán kvasiniek vzorca beta-D-Glcp iYeast wall (1-6) -beta-D-gluco- (1-> 3) -beta-D-glucan of the formula beta-D-Glcp i 6 / \ /l-*3/-beta-D-Gico. í/l-»3/-beta-D-Glcpfc~i^- kde Glcp je glukopyranozylová jednotka. 6 / \ / l- * 3 / -beta-D-Gico. where Glcp is a glucopyranosyl unit.
CS863944A 1986-05-29 1986-05-29 wall / l-> 6 / -beta-o-gluco- / 1 - ► 3 / -beta-o-glucan yeast CS270103B1 (en)

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