CH334922A

CH334922A - Process for the preparation of podophyllotoxin glucoside from podophyllum species

Info

Publication number: CH334922A
Authority: CH
Inventors: Stoll Arthur Dr Prof; Jany Dr Renz; Wartburg Albert Dr Von
Original assignee: Sandoz Ag
Priority date: 1954-10-13
Filing date: 1954-10-13
Publication date: 1958-12-15

Description

  

  



  Verfahren zur Darstellung von   Podophyllotoxinglucosid    aus Podophyllumarten
In der schweizerischen   Hauptpatentsehrift    Nr. 330663 wurde die I) arstellung des bisher unbekannten Podophyllotoxinglucosids aus   Podophyllumarten,    insbesondere aus dem indisehen   Podophyllum    emodi Wall, beschrieben.



  Es wurde nun gefunden, dass dasselbe Glucosid   aueh aus Podophyllum peltatum    L. isoliert werden kann. Die vorliegende Erfindung betrifft ein Verfahren zur Darstellung des freien   mitosehemmenden    und tumorzerstorenden   Podophyllotoxinglucosids    aus   Podophyl-    lum peltatun L., dadurch gekennzeichnet, dass man die Droge mit organischen   Lösungsmit-    teln   auszieht,    den Extrakt konzentriert und dann mit Wasser verd nnt, diese L¯sung zur Trennung des Gerbstoff-Glucosid-Komplexes mit.

     gerbstoffällenden 3fittelll    behandelt, die FÏllung abtrennt, die zur ckbleibende L¯sung von organischen Lösungsmitteln weitgehend befreit und mit Wasser verdünnt, die   wässe-    rige Losung zur Entfernung von Begleitstoffen mit einem mit Wasser nicht mischbaren   Losungsmittel extrahiert, dann    das   Glueosid    mit einem mit Wasser nicht mischbaren L¯sungsmittel, das   höchstens 10"/o eines    wasserl¯slichen aliphatisehen Alkohols enthält, aus  schüttelt, das in    die organische Phase über  gegangene Podophyllotoxinglucosid    nach Ab   destiHieren des Lösungsmittels in fester Form    gewinnt und weiter reinigt.



   Das Podophyllotoxinglueosid hat die   Zu-    sammensetzung C28H32O13 und schmilzt bei   149-152 .    Es zeigt in   Methanollösung die    optische   Drehung      [a]    20/D= -75¯, in Pyridin  l¯sung [&alpha;]20/D= -117¯.    Das   Glucosid    ist in Alkohol und in   Wasser loslieh.    Bei der enzy  matischen    Spaltung und bei der Hydrolyse mit Säure wird das   Glueosid    in Podophyllotoxin und D-Glucose zerlegt.



   Das verfahrensgemäss aus Podophyllumarten dargestellte neue   Glucosid,    das sieh gegenüber den bis jetzt bekannten zuckerfreien Wirkstoffen besonders durch seine Loslichkeit in Wasser auszeichnet, soll für therapeutische Zwecke verwendet werden.



   Beispiel
Getrocknete Rhizome von Podophyllum   peltatum    L. werden fein gemahlen und dann mehrmals bei gewohnlicher Temperatur mit Chloroform extrahiert. Das so vorgereinigte Pflanzenmaterial wird nun mit Methanol ausgezogen, worauf man die vereinigten gelb ge  färbten    Auszüge auf etwa 1/3 konzentriert und dann mit dem gleichen Volumen Wasser versetzt. Die wässerig-methanolische Lösung wird nun zur Trennung des   Gerbstoff-Glucosid-    Komplexes mit einer   konzentrierten wässe-    rigen Lösung von Bleiacetat, solange eine Fällung entsteht, versetzt.

   Nach dem Abfiltrieren des entstandenen   Niedersehlages    befreit man die Losung durch Ronzentrieren im   Vakuurn    mögliehst vollständig vom Methanol, verdünnt mit Wasser und sch ttelt sie mehrmals mit etwas reinem Chloroform aus. Diese Vorfrak tion enthÏlt neben   harzigen    Begleitstoffen auch schon kleine Mengen   Podophyllotoxin-       glucosid.    Die wässerige Restlösung wird an schliessend mehrmals mit Chloroform, dem
10%Alkohol zugesetzt sind, ausgesch ttelt, worauf man die vereinigten fast farblosen
Chloroformauszüge eindampft, den   Eindampf-       rückstand mit. Äther wäscht und trocknet.   



   Die Reinigung dieses   Glucosidpräparates    bereitet ziemliche Schwierigkeiten. Sie geschieht entweder durch Chromatographie an n einer Silicagelsäule aus Essigesterlösung oder    vorteilhafter noeh durch Craigsehe Verteilang    zwischen Wasser und Essigester. Nach beiden Verfahren   erhalt man nach Vorfraktionen, die    hauptsäehlieh noch gefärbte, etwas   harzige    Anteile enthalten, Fraktionen, die aus Podo  phyllotoxinglucosid bestehen.    Das   Glucosid    wird daraus nach Eindampfen und   ITmfällen    aus   Aceton-Äther    in reiner Form gewonnen.



  Dieses Präparat besitzt die oben erwähnten Eigenschaften.



  



  Process for the preparation of podophyllotoxin glucoside from podophyllum species
The preparation of the previously unknown podophyllotoxin glucoside from Podophyllum species, in particular from the Indian Podophyllum emodi Wall, was described in the main Swiss patent document No. 330663.



  It has now been found that the same glucoside can also be isolated from Podophyllum peltatum L. The present invention relates to a method for preparing the free, mitosis-inhibiting and tumor-destroying podophyllotoxin glucoside from Podophyllum peltatun L., characterized in that the drug is extracted with organic solvents, the extract is concentrated and then diluted with water, this solution to separate the tannin-glucoside complex with.

     Treated tannin-precipitating 3fittelll, the filling separated, the remaining solution largely freed of organic solvents and diluted with water, the aqueous solution to remove accompanying substances extracted with a water-immiscible solvent, then the glueoside with a water-not Shake out a miscible solvent that contains a maximum of 10 "/ o of a water-soluble aliphatic alcohol, which, after distilling off the solvent, gains solid form in the organic phase via podophyllotoxin glucoside and purifies it further.



   The podophyllotoxin glueoside has the composition C28H32O13 and melts at 149-152. In methanol solution it shows the optical rotation [a] 20 / D = -75¯, in pyridine solution [α] 20 / D = -117¯. The glucoside can be found in alcohol and water. In enzymatic cleavage and hydrolysis with acid, the glueoside is broken down into podophyllotoxin and D-glucose.



   The new glucoside produced from Podophyllum species according to the method, which is distinguished from the sugar-free active ingredients known up to now especially by its solubility in water, is to be used for therapeutic purposes.



   example
Dried rhizomes of Podophyllum peltatum L. are finely ground and then extracted several times with chloroform at ordinary temperature. The plant material pre-cleaned in this way is then extracted with methanol, whereupon the combined yellow-colored extracts are concentrated to about 1/3 and then the same volume of water is added. To separate the tannin-glucoside complex, the aqueous-methanolic solution is now mixed with a concentrated aqueous solution of lead acetate, as long as a precipitate occurs.

   After the precipitate formed has been filtered off, the solution is freed as completely as possible from methanol by concentrating in vacuo, diluting with water and shaking it out several times with a little pure chloroform. In addition to resinous accompanying substances, this pre-fraction also contains small amounts of podophyllotoxin glucoside. The residual aqueous solution is then several times with chloroform, the
10% alcohol are added, shaken out, whereupon the combined almost colorless
Evaporated chloroform extracts, the evaporation residue with. Ether washes and dries.



   The purification of this glucoside preparation is quite difficult. It is done either by chromatography on a silica gel column from ethyl acetate solution or, more advantageously, by the distribution of water and ethyl acetate. According to both methods, after preliminary fractions, which mainly still contain colored, somewhat resinous components, fractions consisting of podophyllotoxing glucoside are obtained. The glucoside is obtained from it in pure form after evaporation and precipitation from acetone-ether.



  This preparation has the properties mentioned above.

Claims

PATENT CLAIM Process for the preparation of the free mitosis-inhibiting and tumor-destroying podophyllotoxin glucoside from Podophyllum peltatum L., characterized in that the drug is extracted with organic solvents, the extract is concentrated and then diluted with water, this solution for separating the tannin-glucoside complex treated with tanning agents, the precipitate is separated off, the remaining solution is largely freed from organic solvents and diluted with water, the aqueous solution is extracted with a water-immiscible solvent to remove accompanying substances,

Then the lucoside is shaken out with a water-immiscible solvent that contains at most 10% of a water-soluble aliphatic alcohol, which, after the solvent has been distilled off, wins the solid form of the podophyllotoxin glucoside in the organic phase and purifies it further.

Podophyllotoxin glucoside is a white, powdery substance that is soluble in water.

The melting point is 149-152¯; the optical rotation in methanol solution is [a] 20 / D = -75¯. With, ¯-glucosidase the compound is broken down into podophyllotoxin and D-glueose.

SUBMISSION 1. Process according to patent claim, characterized in that, to remove resin substances, the powdered drug is first pre-extracted with a chlorinated hydrocarbon and then the glucoside-tannin complex is extracted with a water-soluble alcohol.

2. The method according to claim, characterized in that the tanning agents from the glueoside-tanning agent complex are precipitated with lead salts and thus the glucoside is released.

3. The method according to claim, characterized in that the drug is extracted with methanol or alcohol and the resin substances are removed by exhaustion with a chlorinated hydrocarbon after the tanning agents have been separated off and before the gucoside is extracted.

4. Method according to claim. characterized in that a mixture of a chlorinated hydrocarbon with a water-soluble alcohol is used to extract the glucoside from the aqueous solution, which is obtained after removing the tannins.

5. The method according to claim, characterized in that acetic acid is used to extract the glueoside from the aqueous solution freed from the tannins.

6. The method according to claim, characterized in that the Podophyllotoxinglucosid dnrell distribution chromatography on a column made of an inert carrier material is further purified.

7. The method according to claim and dependent claim 5, characterized in that the podophyllotoxin glucoside is eluted from the column with ethyl acetate to which a maximum of 5% methanol is added.

S. Verfallren according to claim and dependent claim 5, characterized in that the podophyllotoxin glucoside is eluted from the column with chloroform to which at most 5% methanol is added.

H. The method according to claim, characterized in that the Podophyl- lotoxing glucoside by distribution between water and ethyl acetate, the. a maximum of 10 "/ o methanol are added, subjected to a further cleaning operation.

10. The method according to claim, characterized in that the podophyllotoxin glucoside is further purified by partitioning it between water and chloroform to which at most 10 / o alcohol is added.