CS259571B1

CS259571B1 - Method of bisindole alkaloid insulation

Info

Publication number: CS259571B1
Application number: CS868926A
Authority: CS
Inventors: Eduard Grossmann; Bohumil Proksa; Dusan Uhrin; Zdeno Voticky
Original assignee: Eduard Grossmann; Bohumil Proksa; Dusan Uhrin; Zdeno Voticky
Priority date: 1986-12-04
Filing date: 1986-12-04
Publication date: 1988-10-14
Also published as: CS892686A1

Abstract

Očelom riešenia je izolácia nového bisíndolového alkaloidu vinkarubínu zo zmesi alltaloidov produkovaných rastlinou Vinca minor L. po oddělení vinkamínu. Uvedený účel sa dosiahne extrakciou zmesi alkaloidov nízkopolárnym rozpúšťadlom, protiprúdnou extrakciou zvyšku v systéme benzén— —butanol—kyselina mravčia—tlmivý roztok o pH 3,5 a chromatografickým přečištěním surového vinkarubínu na oxide hlinitom. Pri testoch na buňky leukémie P-388 vinkarubín preukázal potenciálně protinádorové účinky a preto bude mať použitie vo farmaceutlckom priemysle.The solution is to isolate the new bisindole vincarubine alkaloid from the mixture alltaloids produced by Vinca minor L. after vincamine separation. stated the purpose is achieved by extraction of the alkaloid mixture low polarity solvent, countercurrent extraction of the residue in the benzene system - —Butanol — formic acid — buffer pH 3.5 and chromatographic purification of crude vincarubin on alumina. In P-388 leukemia cell assays, vincarubin potentially anticancer effects and therefore will be of use in pharmaceuticals industry.

Description

253571

The present invention relates to a process for the isolation of a bisindole alkaloid of the vincarubin of formula I from a mixture of alkaloids produced by Vinca minor L., after separation of the vincamine.

From the mixture of alkaloids produced by Vinca minor L., vincamine, which has been introduced into the practice of medicine as a low-toxicity cerebrospinal fluid and hypotensive, is isolated in the industrial plant. Vincarubin, the first bisindole alkaloid isolated from the plant, is a novel, not yet described substance, characterized by potential antitumor effects. In evaluating the effect of vincarubin on leukemiaP-388 cells, the greater activity of this substance (Table 1) was shown as vinblastine, a bisindole al-caloid isolated from a Catharanthusroseus plant used in clinical ipraxi.

In the process of isolating the vincarubin according to the invention, the mixture of alkaloids isolated from the Vinca minor L. plant after the separation of the vincamine is extracted with a low polar solvent such as diethyl ether or dichloromethane. the extract is concentrated, subjected to countercurrent extraction in benzene-butanol-formic acid - pH 3.5 buffer (25: 5: 1: 31), the vincarubin-containing solution being concentrated and purified by column chromatography on oxide aluminum in a system consisting of an aromatic hydrocarbon and an alcohol having carbon numbers of C1 or C2, for example benzene-methanol (9: 1) or toluene-ethanol (9: 1).

Further details of the method of isolation of the invention are apparent from the example of conversion. Example 1

The dry drug (300 kg) from Vindaminor L. was mixed with 10% aqueous ammonium hydroxide (240 L), after 24 h the sasuspension was extracted 5 times after 500 l benzene. The benzene extract was concentrated, the residue triturated 6 times with 15 1 of 0.5 ° / hydrochloric acid. The combined acidic extracts were adjusted to pH 8.5 by the addition of ammonium hydroxide, extracted 5 times with 10 L of chloroform. The chloroform extract was concentrated and the residue was separated by column chromatography on alumina (12 kg) eluting with diethyl ether (160 L) and methanol (10 L). 50 g of (+) - vincamine were obtained from the concentrated ethereal eluate by crystallization. Example 2

The mixture of substances (1 kg) which remained after the separation of (+) - vincamine was triturated with 5-fold in 6 l of diethyl ether (or dichloromethane), the extract was concentrated (residue 75 g) and subjected to countercurrent extraction in the benzene-butanol system formic acid buffer opH 3.5 (Mc Ilvain solution) (25: 5: 1: 31), the vincarubin-containing portion (appearing in red) was separated and concentrated. 7.7 g of crude vincarbine were obtained, which was purified by column chromatography on alumina (300 g) in the form of benzene-methanol (9: 1) or toluene-ethanol (9: 1).

Concentration of the vincarbine-containing fraction gave pure material, C 43 H 50 N 4 O 6 (718.9), mp 170 ° C (decomposition),

Λ max (log ε, mzm -1), nm: 273 (3.21), 340 (3.29), 480 (2.40), IC (KBr): 1,730, 1,680, 1,605 cm -1, [α] D 23 -550 + 10 ° (c 0.1 ethanol),

Rf = 0.5 (silica gel, benzene-ethyl acetate-ethanol 20: 29: 1).

Claims

OBJECT OF THE INVENTION

A process for the isolation of a vinca-carbine bisindole alkaloid of formula I (1) from a mixture of alkaloids produced by Vinca minor L. after separation of vincamine, characterized in that the alkaloid mixture is extracted with a low polar solvent, the extract is concentrated. butanol-formic acid buffer, pH 3.5, the vincarubin-containing fraction is concentrated and purified by chromatography on alumina in a system consisting of an aromatic hydrocarbon and an alcohol having a carbon number of C1 or C2.

2. The method according to claim 1, wherein the low-polar solvent is diethyl ether or dichloromethane.

3. The method of claim 1, wherein the countercurrent extraction is carried out in a benzene-butanol-formic acid-buffer system at pH 3.5, at a volume ratio of 25: 5: 1: 31 by volume of the components.

4. The method according to claim 1, characterized in that benzene or toluene is used as the aromatic hydrocarbon for the chromatography.