DE1593086A1 - Process for the production of Peruvoside - Google Patents

Description

(previously; M 70 969 IVb / 12ο)

E. M e r c k

Corporation

Darmstadt 1593086 ⁷ * ^{September iQbb}

Process for the production of Peruvoside

£ as, .Crlukosid "Peruvosid" has a pronounced cardioto-. nisohe effect, which roughly corresponds to that of strophantine. In contrast to this therapeutically widely used gardenolide, however, it is absorbed considerably better when administered orally. Peruvosid is a natural product and, for example, in the Apocynacee Thevetia neriifolia Juss. (syn. Thevetia peruviana Merr.) included. It is known to isolate the compound from the defatted and fermented seeds of these plants by extraction and chromatographic separation of the extracted substances in Helvetica Qhimiea Aota, Ba »4 43, page 652 Ci9fe ©} and volume 45» page 958 (1962). In these known "isolation" and purification processes γ / earth alcohols are used as solvents. Me yields of Peruvoside in the known processes are extremely low and in most cases not reproducible.

It has been found * that, surprisingly, considerably higher and clearly reproducible yields are obtained if "the isolation of the peruvoside with organic solvents which have no active hydrogen" is carried out.

Subject dter Yo ^ i «« «» Direction> invention accordingly hite method * aur isolation of Peruvosid from defatted fermented "peruvofiidhaltigem plant material, in particular seeds of fhevetia" Erii folia Juss, by extraction followed chromatographißchö separation ®nü the extracted substances "( Yes ■ is characterized by "**» ** that: all stages of the process are carried out with organic solution custom. They do not have any active »// asaer & to-ff»

BATH ORIGINAL

ils solvents that do not contain active hydrogen come For example, the following in embossing χ halogenated hydrocarbons, / ie chloroform, methylene chloride, 1,1-dichloroethane, 1,2-diehlorethane, , 1,2-trichloroethane, 1, i-dichloro-Z-Iluor-ethane, carboxylic acid ester, ie methyl acetate, ethyl acetate, ethyl formate ster, propionic acid methyl ester or oyclieche ethers »like dioxane, tetrahydrofuran or ketones such as acetone, ethyl methyl ketone or diethy1sulfoxide or dimethylformamide and others polar organic solvents that do not have any active hydrogen in the molecule; The preferred solvent for extraction is chloroform.

oh the new method according to the present invention, rhält Peruvosid in yields of about 1 to 1.2 i "based on as defatted unfermented starting material · The yield achieved so st, for example, twice as high as that according to the method, it Indian patents 66,196 yield reached "

The new procedure is carried out individually as follows »

as used as starting material defatted fermented _t peruosidhaltige plant material is from dexjzerkleinerten peruvoaidaltigen parts of plants, preferably obtained from seeds of Thevetia neriiolia in a known manner · The following procedure has proved expedient for he preparation of the starting material:

Initially, the glycoside-containing plant material is squeezed and then greased with a fat-dissolving organic solvent, for example with a hydrocarbon such as petroleum ether, a halocarbon such as chloroform or an ether such as diethyl ether. The pre-degreased material is then ground again ad dried again with a fat-dissolving organic solvent _e

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ft OPO 1 f + π ** tn

i> er β ο. The finely ground tiücksland obtained is fermented in the usual way in aqueous suspension. A small amount of toluene is advantageously added to the fermentation mixture in order to prevent spoilage of the plant material due to the action of microorganisms such as bacteria and fungi. The fermentation time is generally between 3 and 5 days, preferably around 3 days 5O ⁰ C, preferably at about 40 ⁰ G ₀ .For use for the method according to the present invention is advantageously the Nasser from the fermented plant material removed by centrifuging

In the '1st stage of the process according to the invention, the fermented, degreased starting material with an organic solvent which has no active hydrogen, preferably with Chloroform, extracted.

By adding the extract, the Peruvoside is obtained in a mixture with other substances. If defatted fermented seeds of Thevetia neriifolia are used as the starting material, when the extract is concentrated, a crystalline mixture of about a rope of Peruvoside with about 2 parts of excess molten falls with it smaller additions of euvoside from _o

Organic solvents with active substances are unsuitable for extraction. Hydrogen in the molecule, which is produced, for example, according to the method of Zerewitinoff (reports of the German onemic society, volume Page 2023 (1907)) can be determined «, For example, connections with OH groups, especially alcohols, and also compounds with ADino, carboxy and / or mercapto groups not for extraction suitable. On the other hand, the presence of drums reduces during the extraction the yields do not.

The separation of the extraction mixture in the 2 _O otufe of the inventive method is carried out according to the usual methods of A-dsaeptionschromatographie. Ken can chromatographically separate the mixture in a column or in a layer

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the usual chromatography compounds in embossing ,, for example finely powdered silica gel, aluminum oxide, magnesium ailicate or Calcium phosphates. The preferred sorbent is silica gel.

By using suitable eluents or hotmelts, for example the above-mentioned solvents, a separation is achieved of the desired pure Peruvosid from the undesirable accompanying substances like neriifolin. £ s has proven particularly beneficial for ils the chromatograph! see separation use the same solvent, which is also used for the previous extraction of the JPeruvoside was, especially chloroform · In a preferred embodiment of the process, the mixture obtained after the first process stage is aue Peruvosid with the undesired accompanying substances, for example neriifolin, by elution with a halocarbon * hydrogen, especially chloroform, over 8-10 times the amount Sorbent (preferably silica gel), bezotjen on the amount of Mixtures to be separated, chromatographed. In this embodiment For example, from the above mixture of 1 part Peruvosid with 2 parts Neriifolin the üeriifolin is practically quantitative first eluted. This is followed during the elution by a small mixture zone of iseriifolin and peruvoside and finally a fraction which consists of pure Peruvoside.

In some cases it is useful to do this after the chromatographic Separation accruing Peruvoeid to recrystallize again. for one Any further purification that may be required, it is advantageous to eat, also an organic solvent which is not active Contains hydrogen, use. For example, the extraction and chromatographic recrystallization is also suitable Separation preferably used chloroform. >

s new process according to the present invention explained in more detail by the following examples:

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009831/1789 * " ^0WÖWAL

example 1

2kg peeled Müsse of Thevetia neriifolia be squeezed and degreased with chloroform, ftach grinding and degreasing again, with chloroform, the residue teigt with water and fermented for 70 hours at 40 ⁰ C with the addition of about 10 ml toluene, The ur> eh moist drug is then extracted exhaustively with chloroform and the extract obtained is concentrated to approx. 200 ml. 35 to 40 g of a mixture of Peruvoside and ISeriifolin crystallize out.

The mixture obtained in this way is freed from the mother liquor and separated on 400 g of silica gel for chromatography (0.05 »0.2 m particle size) with chloroform as the eluent. The first fraction consists of approx. 23 to 27 g of pure neriifolin, followed by an intermediate fraction of 1 to 2 g of a mixture of Peruvoside / Jieriifolin and a last fraction, which consists of 11 to 12 g of pure Peruvoside. The entire amount of Peruvoside is dissolved in 6 ml of acetone and 6 ml of diethyl ether are added. The Peruvosid crystallizes in prisms or druses. The yield is 10 to 11 g. The melting point is between 160 and 163 °. C ^ J ψ ^s ~ ⁷⁰ ° Z ² ° (c ^{β 1} * 3 in methanol). The substance is pure according to thin-layer chromatography.

Example 2

The chloroxorm extract prepared according to Example 1 of the fermented The seeds of l'hevetia neriifolia are mixed with a conventional desiccant dried like sodium sulfate and treated with diethyl ether. Included a crystalline mixture of Peruvosid, Neriifolin and ituvo & id separates out, which after filtering off and drying has a weight of has about 50 g.

Fitian chromatographed the mixture obtained over 350 g of silica gel analogously to Example 1 with methylene chloride as a cluting agent. The following fractions are then in the following iieihenfolge eluierti Ca ₀ 25 g of pure Neriifolin, about 4 g of Übergangfjfraktion lieriifolin and Peruvosid, about 11 g reinee Peruvosid and finally., 3?. a mixture of 2 parts Peruvoaid and one part Ruvosid. The Peruvosid is as described in Example 1, recrystallized and has the physical properties specified there. "Γ7Γ ~ * ~

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009831/1789 _Ä

Claims (1)

Fa tentana 1. Process for the isolation of Peruvoside from defatted, fermented, peruvoside-containing plant materials, in particular Seeds of ^ hevetia neriifolia Jmss. (syn. Thevetia peruviana Herr «), by extraction and subsequent chromatographic separation of the extracted substances, characterized in that both stages of the process are carried out with organic solvents which do not contain active hydrogen » 2 * Method according to claim 1, characterized »that for the Extraction and the same solvent, preferably chloroform, is used for chromatographic separation » 5 «Method according to claim 1 and 2, characterized» that one the chromatographic separation at the 6 to 12-fold amount Sorbent, preferably 8 to 10 times the amount of silica gel, based on the weight of the su separating substances. New subject <* t 1 1 «** = * ■ ι * .u 3 *. * n * ™ «- * 4. β. ικ » BAD ORiGJNAL 009831/1789