IL34740A - Sulfonated derivatives of a glycopeptide extracted from animal organs and their preparation - Google Patents

Description

derivatives of a extracted animal their preparation 35027 drosses by with an enzyme papain or in a proper pH to It can also be convenient to before the enzymatic a preliminary precipitation of most of the proteic drosses by adding a suitable such for acetic By diluting the filtrates obtained from these with organic solvents or the raw giycopeptide is obtained in the form of a white to hazel The products contain only a trace of sulfur and characterized by the absence of amines are present in the range of amino acids and The above analytical data permits that the products be classed among the on the other the presence of hexosaminea accompanied by acids and the absence of groupings o all confer a characteristic feature upon the polysaccharide moiety of this The giycopeptide obtained as described in cation Serial is suspended in a heterocyclic tertiary base having a boiling point of from about to about Typical bases include methyl ethyl and dimethyl Anhydrous pyridine is The giycopeptides are insoluble i the latter such that they are in suspension Suitable sulfonating agents include chlorosulfonia sulfuric oleum and adducts of sulfuric anhydride Organic compounds such as pyridine and Sulfonatio is effected by bringing the tide suspension and the sulfonating together at a eraturQ from about to then the same at a temperature from about to about and preferably at abou for from about 4 to about and preferably The quantity of sulfonating agent can b varied from about 400 to about and ably about percent by weight the At the end of the sulfonation the fonate product is collected by filtration and the impurities of basic or acidic nature are removed by or by treatment with ionic exchange SULFONATE SALTS The sulfonat glycopeptid hereinafter be also indicated as is then converted to a sodium salt with sodium and the sodium salt is precipitated by dilution of its water solution with able as acetone or another aspect of the the SGLP is converted t other salts other metals or or or heavy The salts of the SGLP are obtained the same SGLP originating front aulfonation through neutralisation with an alkaline or metal hydroxide or ammonium as well as the SGLP sodium salt by double exchange with an o heavy metal Salts obtained through SGLP neutralization with metal hydroxide are isolated by th aqueous solution with a precipitating agent as acetone or has been found that particularly favorable results are obtained when before the precipitation a small amount o from 3 to about 7 percent b based upon the weight of the is added of an acetate or of the same metal whose hydroxide was employed for which are most suitable embodiment are potassium strontium and Double exchange reaction between SGLP sodium salt and salts of other metals carried out in aqueous the anion of the salt under consideration being stituted preferably of an acetate or of a exchange reaction ma be carried out also by percolating an aqueous solution o sodium salt through a column of strongly cationic exchange resin which was previously with the metal whose salt is to be The percolated solution is then treated with a precipitating as acetone or preferably after addition of a amount of a acetate or h lide of the meta whos salt is desired as mentioned particularl suitable for this double exchange reaction are the following gold and In the following Table some analytical values reported which are characteristic for the sodium salt of SGLP prepared as corresponding to materials with the lowest and highest sulfonatlon degree S C H β 0 Hexosamines 14 CH3CO Proteins acids The proteic component has the following composition percentage Threonine Serine Glutamic Alanine Valine Leucine Lysine Hystidlne In the tests On laboratory th glycopeptide sulfonated derivatives have proved to be active in the inhibition the local oedema in the paw caused by serotonine and the acitivity shows itself alread at inhibition off wheals at endope increase off the functional recovery rate of the limb with arthritis by at increase of the rate of mental wounds at and at kg subcutaneously inhibition of the ulcer by pylorus ligature and by at mgAg peritoneally and at 50 mg kg decrease of the peptic acitivity of the gastric juice either or increase of the fibrinolytic acitivity as determined by the methods of the lysis of the plates according to Astrup and said activity shows itself at 500 gamma decrease the cholesterol and of plasmatic after treatment in the NO toxicity proof was evidenced in rats and after single treatment with 3 gAg or prolonged administration for 6 months 100 following examples illustrate the although they have no limiting character 1 One gram of extractive glycopeptide obtained as described in Example 5 of application go filed on September is suspended in 20 ml anhydrous The solution is cooled while stirring at and is treated slowly with g of chlorosulfonic Afte carrying out the the reactio is completed by heating for 6 hours at then allowed to cool and resulting precipitate is collected by filtratio and then washed with The product is afterwards solved in 50 ml water the solution is to pH 10 by adding 1 normal filtered the of the filtrate is adjusted to 6 by adding acetic the solution is 48 distilled at room Wit 7a regenerated cellulose the end of the dialysis oper solution is and th is Yield s Analysis S acids Acetyl groups Proteins Example 2 One gram of the glycopeptide of Israel specification in Example suspended in 20 ml of anhydrous The suspension is cooled at while it is stirred and slowly treated with g of chlorosulfonic After carrying out the the reaction is brought pletion by heating the reaction mixture for 6 hours at then it is allowed to and the resulting precipitate is collected by The precipitate is dissolved in 50 the is precipitated by diluting with methanol and collected again by The precipitate is dissolved more in water the solution is treated whiie for half an with a mixture of ion exchange resins consisting o 25 120 resin and 25 ml o IRA 410 The resins ar removed by the adjusted to 6 by addition normal and the solution is concentrated unde reduced pressure to a volume 15 The product is precipitated by dilution with 60 ml methanol adding to the solutio g hydrated sodium The precipitate is collected on a washed with with ether and dried in vacuum upon Na acids Acetyl groups Proteins 2 g of glycope mentioned Patent Specification 28649 I Example suspended in 40 ml of anhydrous The suspension is cooled at and treated slowly whil stirred with of chlorosulfonic After carrying out the the reaction is brought to completion by for one hour stirring and for two hours without stirring at a temperature of The solvent is decanted and the resulting is taken up again with th collected on The product is dried unde at dissolved in water and treated while stirring with 25 of ion exchange resin 120 for 30 The resin is removed by filtration and filtrat undergoes the treatment with 25 ml of exchange resin The latter is removed by filtration and the pH of the tion is adjusted to 11 b 1 normal The filtrate brought to pH by treating it with 2 then g of hydrated sodium acetate is the liquor is concentrated under reduced pressure a of 20 ml and the product is precipitated b dilution with 80 of The product is collected on with alcohol and then dried in vacuum at Yields 3 S Eexoses Uronic acids Acetyl groups Proteins Example 4 An extractive glycopeptide is subjected to tion according to one of the Examples An aqueous solution of acidic as it is obtained out the reaction is adjusted to H with ba hydro Then it is under reduced pressure a volume of 15 g of is added and it is diluted again with 60 of methanol to obtain a precipitate constituted of the fonated glycopeptide barium E ample 10 g a glycopeptide sodium salt obtained as described in Example are dissolve in 100 ml of water and treated with an aqueous solution 1 zinc it is diluted with 400 methanol to obtain a precipitate constituted of the ated glycopeptide zinc Zn S 55 ml of a strongly ic exchange A EE has bee activated b known is charged to an 80 high glass wherethrough one liter of is percolated within about one The column is subsequently thoroughly washed b allowing 500 ml distilled water to flow therethrough within an Through the said a solution is then passed which contains 20 g of sulfonated glycopeptide sodium salt as in S in 800 ml of distilled water within about The column the with 500 mi of distilled then the washing waters and previously obtained eluates are The solution is concentrated under reduced pressure to a volume of 600 g are added and is diluted with 1200 ml of acetone to obtain precipitate constituted of the sulfonated peptide aluminum Analysisj S 7 100 g of a sodium salt of glycopeptide sulfonate as in were dissolved in 1000 ml of lukewarm water and poured 1000 mi of a molar bismuth nitrate solutio in glacial acetic Then 2000 ml of methanol were The resulting precipitate was and washed three by crushin in 1500 ml of acetic acid mixture and it each Then the precipitate washed again three times i the sam manner using 1500 ml of acetone each The powder which obtained was dried in and j dissolved again 15 volumes of wate to obtain a acid solution An aqueous 1 normal caustic soda solution was added to provide a solution having a pH and then a as obtained by adding volumes of acetone to the precipitate was centrifuged and washed three times with a it each and then it was washed three times with the precipitate was dried in vacuo to give 120 of 3 A gold salt corresponding to the bismuth 7 was prepared by the procedure of the latter with the exception of using a auric nitrate solution in glacial acetic By proceeding analogously to the procedures cribed in illustrative the following are Magnesium salt of the sulfonated glycopeptid S Cobalt salt of the sulfonated Co S salt of the sulfonated Ni S Copper salt of the sulfonated glycopeptid Cu S Calcium salt of the sulfonated Ca S insufficientOCRQuality

Claims (1)

1. WE CLAIM IS i A process for preparing a sulfonated produc of a glycopeptide from the gastric mucosa or duodenum of hydrolyzing the animal organ in at a temperature from about to abou for from about 10 minutes to about 45 minutes at a from about 1 to removing acidic hydrolysis obtained by the hydrolysis of diluting the product of with a therefor whereby a glycopeptide is precipitated suspending the product of in a cyclic tertiary base having a boiling point of from about to about contacting the resulting suspension of with a sulfonating agent selected the group consisting of sulfuric acid and an adduct of anhydride an organic at temperature from about to about then a temperature from about to about and recovering the resulting fonated The process of Claim 1 wherein the base is The process of claim 1 wherein the sulfonating agent of is chlorosulfonic The process of Claim 1 wherein the contac time with the sulfonating i is from about 4 t about 8 The process of Claim 1 wherein the sulfonated product formed in is The process of Claim 1 wherein the product formed in is contacted with an ion exchange resin selected from the group consisting of an anionic a c and a mixture of an anionic and a cationic exchange The process of 1 wherein the sulfonated product recovered in with a hydroxide selected from the group consisting of an alkali metal an alkaline earth metal hydroxide and ammonium by sulfonated product is converted to its corresponding alkaline earth metal or ammonium and the suiting product with acetone o methyl alcohol whereby said salt is The process of Claim 7 wherein a mino amount of an acetate or halide having the same cation as said hydroxide is dissolved i the acetone or methyl The process of Claim 7 wherein the hydroxide of is sodium hydroxide and the salt of is sodium The process Claim 9 wherein a solution of the said sodium salt is treated with a halide or acetate a metal from the group consisting of aluminum and and the resulting metal salt with acetone or methyl The process of Claim wherein an aqueous solution of the said sodium salt is percolated through a cationic exchange resin having as a cation a metal selected from the group consisting of and contacting the resulting percolated solution with a minor amount of an acetate or halide of and precipitating the resulting metal salt with acetone or methyl The process of Claim 1 wherein the of comprises from about to about 3 volumes methyl alcohol or ethyl fo each product of process Claim 11 wherein the of acetate or halide is from about 3 to about 7 percent by a sulfonated glycopeptide 1 th cation of which is selected the grou consisting of an an alkaline earth heavy and A salt of Claim 14 wherein the cation A sodium salt of Claim 15 having the ing approximate composition in percent by S H 0 Hexosamines Hexoses roteins and being substantially of phosphorus and uronic A salt of 1 wherein the cation is A salt of Claim 14 wherein the catio is A salt of Claim 14 Wherein cation is A t of Claim 14 wherein the catio is A salt of Claim 14 wherein the cation is A salt of 14 wherein the catio is A salt of Claim 14 wherein the cation is A salt o Claim 14 the cation is A sal of Claim wherein the cation is A salt of Claim 14 wherein the cation is insufficientOCRQuality