CH166354A - Method for the preparation of a cardiac active glucoside. - Google Patents

Description

  

  Method for the preparation of a drug-active glycoside. WA Jacobs and A. Hoffmann have shown (Journ. Biol. Chemistry 69, 153-163, 1926, and Chem. Zentralblatt 1927, I, page 294) that it is possible to convert K-strophantine into cyrnarin and glucose with the help of a ferment to split. The ferment (strophantobiase) only splits off the glucose of strophantine and is strictly specific.

   It was therefore not foreseeable that other drugs containing cardiac glucosides could contain fer ments that would split off part of the sugar from glucosides.



  It has now been found that in the digitalis species (such as, for example, Digitalis purea, Digitalis lanata rrsw.) And in the sea onion (Scilla maritima), enzymes are found which partially hydrolyze the cardiac glucosides of these drugs and, under suitable conditions, a part of the sugar of the glucoside molecules are able to split off.



  The present invention relates to the partial splitting off of sugar from glucoside molecules with the aid of ferments present in the drug. It makes it possible to convert heart glucosides that are richer in sugar into glucose-free glucosides, be it by allowing the process to take place during the extraction or even before it under suitable conditions, or by starting from pure glucosides that have already been isolated in a mixture or from pure individual individuals .

      If you want to split the glucose before extracting the glucoside from the drug, it is advisable to let the sea onion stand for some time with water and an extractant. Since the enzymes contained in the drug split the (älucose of the rope A, so that the Proseillaridin A is obtained during the extraction.



  The subject of the present patent is a process for the preparation of procillaridin A, which is characterized in that scillaries A are subjected to the action of a sugar-splitting enzyme from the sea onion. To carry out this process, pure or fiber-containing rope A or sea onion, which contains rope A, can be used as the starting product.



  <I>. </I> Example <I> 1: </I> 10 kg of fresh sea onions are cut into coarse pieces and stored under ethyl acetate for 5 days, the action of the ethyl acetate killing the living cells of the fresh drug, without a putrefaction process occurring.

   On the other hand, in this treatment the enzyme specific for rope A, the scillarenase, unfolds its effect by partial hydrolysis of the rope A contained in the sea onion, which is made up of its aglycon scillaridirr A, rhamnose and glucose, by partial hydrolysis into that poor in glucose Glucoside, the proscillaridin A, transferred. Then the ethyl acetate is poured off and the onion pieces are crushed together with 10 kg of ammonium sulfate to a fine paste, pressed out, and the residue is exhaustively extracted with ethyl acetate.

   The ethyl acetate poured off at the top can also be used here. The acetic ester extract is completely evaporated in vacuo, the residue washed with ether several times or. boiled, the remaining undissolved portion dissolved in one liter of a mixture of methanol-water 1: 1, treated with an insoluble tannin precipitant, filtered and the solution freed from methanol in vacuo.

   Proscillaridin A separates out in crude form, is filtered, precipitated from methyl alcohol by adding water and then recrystallized from methyl alcohol.



  The hitherto unknown proscillaridin A is composed of a molecule of scillaridin A and a molecule of rhamnose and is easily split into these by hydrolysis with acid at the split pieces, both of which are obtained in crystallized form.



       Proscillaridin A crystallizes from methyl alcohol in thick plates which contain crystal solvents and which weather when standing in the air or in a desiccator. It dissolves in around 25 parts of methyl alcohol, around 50 parts of alcohol and 600 parts of chloroform, and is extremely sparingly soluble in water and ether. In 80% alcohol it has a spec. opt. Rotational power of [a] D - - 860 (20) (c - 2.4), based on the dried substance.



  Like Seillaren A and Scillaridin A, Proscillaridin A also produces the Liebermann cholestol reaction with acetic anhydride and H280.



  <I> Example 2: </I> 3 gr ropes A are finely ground, intimately mixed with 70 gr of the enzyme preparation prepared as described below, 200 cubic meters of water, 2.5 cubic centimeters of acetic acid 1:10 and 5 cm of toluene and left to stand at 360 ° C. for 20 hours with frequent stirring. It is then filtered, washed with water until the reaction is neutral and the residue is exhaustively extracted with alcohol, the degraded glucoside being dissolved.

   The alcoholic solution is completely evaporated in vacuo, the residue is taken up in about 11/2 liters of chloroform, the proscillaridin A dissolving. The filtered solution is evaporated in vacuo and the residue is first precipitated from methanol by the addition of water, filtered and then recrystallized from methyl alcohol. The preparation shows all of the properties of proscillaridine A given in Example 1.



  The enzyme preparation used in the present example was presented as follows: 2 kg of fresh sea onions are chopped up and left to stand for 14 days with 8 liters of water which is saturated with thymol, during which autolysis occurs. Then it is rubbed through a cloth and squeezed out. The golat is filtered through a talc filter, the clogging layer that adheres to the talc surface is scraped off from time to time. The scraped-off material contains the active enzyme in a highly concentrated form. It is collected, washed first with water, then with alcohol and quickly dried.

 

Claims (1)

PATENT CLAIM: Process for the preparation of proscillari- din A, characterized in that ropes A are subjected to the action of a sugar-splitting enzyme from the sea onion. Proscillaridin A is made up of a molecule of scillaridin A and a molecule of rhamnose and is easily split into these two fragments by hydrolysis with acid, both of which are obtained in crystallized form. Proscillaridin A crystallizes from methyl alcohol in thick plates that contain crystal solvents and weather when standing in the air or in a desiccator. It dissolves in around 25 parts of methyl alcohol, around <B> 50 </B> parts of alcohol and 600 parts of chloroform, and is extremely sparingly soluble in water and ether. It has a spec in 80% alcohol. opt. Rotational power of [a] D = - <B> 861 </B> (-f- 20) (c = 2.4), based on the dried substance. Like Seillaren A and Scillaridin A, Proscillaridin A also produces the Liebermann cholestol reaction with acetic anhydride and H2S04. Proscillaridin A is said to be used in therapy. SUBClaims: 1. Method according to claim, characterized in that sea onion is left to stand with water and an extractant until the glucose is split off from the rope A. 2. A method according to claim, characterized in that sea onion is brought together with a medium which induces cell death and at the same time prevents putrefaction and, after glucose has been split off from rope A, the scillaridin A is extracted from the reaction mixture. 3. The method according to claim, characterized in that pure rope A is used as the starting product and this is subjected to the action of an enzyme preparation obtained from the sea onion.