Anti-tumour effective constituent cherimoya essence preparation method

Abstract

The invention provides a process for preparing Fanlixin, an active antineoplastic ingredient in custard apple from custard apple seeds as raw material through organic solvent extraction, abstraction, preparing custard apple total lactone, refined isolation through column chromatography and / or reversed phase C18filling column. The Fanlixin content amounts to above 90.0%.

Description

Technical field

[0001] The present invention belongs to the preparation method of plant active ingredient, mainly relates to the preparation method of a kind of annonaceous lactone (Annonaceousacetogenins) Annonaceae (Annonaceae) Annonaceae (Annonaceae) plant Annona squamosa L. Annona cinnamon is an effective antitumor ingredient. Background technique

[0002] The custard apple is a kind of fruit tree introduced and cultivated in tropical and subtropical regions of my country. It is native to tropical America. The fruit is edible and sweet. Its seeds contain anemone lactones. At present, nearly 20 such compounds have been isolated from custard apple plants. Most of these natural anemone lactones have different degrees of antitumor activity. Its molecular structure has a long-chain fatty acid basic skeleton, an α, β unsaturated γ-lactone ring at the end, 0-3 tetrahydrofuran rings and several oxygen-containing substituents in the middle of the long chain, and there are more...

Examples

Embodiment 1

[0020] Preparation of total lactones: get 5 kg of custard apple seeds, grind into coarse powder, in the pulverizing process, remove seed shell as far as possible, soak seed coarse powder in water for 24 hours, filter off water, wash with water, after centrifugal dehydration, infiltrate with dehydrated alcohol Extract by filtration, recover ethanol to liquid extract at 42°C, add 1.5 liters of 15% ethanol to suspend, degrease with petroleum ether, 4000ml each time, three times in total, combine petroleum ether extracts, concentrate to 3000ml, and extract with 1000ml 80% ethanol for 3 The second time, combine the 80% ethanol extract, recover it under reduced pressure at 42°C to about 1000ml, combine it with the dilute alcohol liquid extracted by petroleum ether, remove the petroleum ether therein under reduced pressure at 42°C, and cool down when the vacuum degree rises to 0.09Mpa. Bring to room temperature, adjust the alcohol content to 15%, extract four times with chloroform, 40...

Embodiment 2

[0022] Take 0.5 kg of custard apple seed shell, extract by percolation with absolute ethanol, reclaim ethanol to form liquid extract, suspend with 15% ethanol, extract three times with petroleum ether, 200ml each time, recover under reduced pressure to remove the dilute alcohol solution. After petroleum ether, it was extracted with chloroform, and the chloroform was recovered to obtain 24.5 g of extract, which was determined by thin-layer chromatography and high-performance liquid phase, and the content of active ingredients was very small. In view of the low content of active ingredients in the seed shell, it can be removed.

Embodiment 3

[0024] Prepare the total lactones according to the implementation case 2, obtain 117.2g total lactone extracts from 5kg raw materials, get 30g total lactone samples, dissolve the sample with petroleum ether-ethyl acetate (7: 3) 200ml, pass through 300-400 purpose Silica gel column pressurized separation, the inner diameter of the silica gel column x length is 70 x 460mm, initially 9000ml was eluted with petroleum ether-ethyl acetate (7:3), and then eluted with petroleum ether-ethyl acetate (3:7). When the eluent is pale and no desired components are eluted, concentrate the 3:7 eluent to dry extract, dissolve it with 500ml of methanol-water (76:24), and use C 18 Packed column pressurized separation, C 18 Packing column inner diameter x length is 70 x 920mm, C 18 The particle size of the filler is 40-63 μm, eluted with methanol-water (76:24), the detection wavelength is 210nm, the required fraction is collected, the solvent is removed below 42°C, and further freeze-dried to obt...

Technical field

[0001] The present invention belongs to the preparation method of plant active ingredient, mainly relates to the preparation method of a kind of annonaceous lactone (Annonaceousacetogenins) Annonaceae (Annonaceae) Annonaceae (Annonaceae) plant Annona squamosa L. Annona cinnamon is an effective antitumor ingredient. Background technique

[0002] The custard apple is a kind of fruit tree introduced and cultivated in tropical and subtropical regions of my country. It is native to tropical America. The fruit is edible and sweet. Its seeds contain anemone lactones. At present, nearly 20 such compounds have been isolated from custard apple plants. Most of these natural anemone lactones have different degrees of antitumor activity. Its molecular structure has a long-chain fatty acid basic skeleton, an α, β unsaturated γ-lactone ring at the end, 0-3 tetrahydrofuran rings and several oxygen-containing substituents in the middle of the long chain, and there are more...

Examples

Embodiment 1

[0020] Preparation of total lactones: get 5 kg of custard apple seeds, grind into coarse powder, in the pulverizing process, remove seed shell as far as possible, soak seed coarse powder in water for 24 hours, filter off water, wash with water, after centrifugal dehydration, infiltrate with dehydrated alcohol Extract by filtration, recover ethanol to liquid extract at 42°C, add 1.5 liters of 15% ethanol to suspend, degrease with petroleum ether, 4000ml each time, three times in total, combine petroleum ether extracts, concentrate to 3000ml, and extract with 1000ml 80% ethanol for 3 The second time, combine the 80% ethanol extract, recover it under reduced pressure at 42°C to about 1000ml, combine it with the dilute alcohol liquid extracted by petroleum ether, remove the petroleum ether therein under reduced pressure at 42°C, and cool down when the vacuum degree rises to 0.09Mpa. Bring to room temperature, adjust the alcohol content to 15%, extract four times with chloroform, 40...

Embodiment 2

[0022] Take 0.5 kg of custard apple seed shell, extract by percolation with absolute ethanol, reclaim ethanol to form liquid extract, suspend with 15% ethanol, extract three times with petroleum ether, 200ml each time, recover under reduced pressure to remove the dilute alcohol solution. After petroleum ether, it was extracted with chloroform, and the chloroform was recovered to obtain 24.5 g of extract, which was determined by thin-layer chromatography and high-performance liquid phase, and the content of active ingredients was very small. In view of the low content of active ingredients in the seed shell, it can be removed.

Embodiment 3

[0024] Prepare the total lactones according to the implementation case 2, obtain 117.2g total lactone extracts from 5kg raw materials, get 30g total lactone samples, dissolve the sample with petroleum ether-ethyl acetate (7: 3) 200ml, pass through 300-400 purpose Silica gel column pressurized separation, the inner diameter of the silica gel column x length is 70 x 460mm, initially 9000ml was eluted with petroleum ether-ethyl acetate (7:3), and then eluted with petroleum ether-ethyl acetate (3:7). When the eluent is pale and no desired components are eluted, concentrate the 3:7 eluent to dry extract, dissolve it with 500ml of methanol-water (76:24), and use C 18 Packed column pressurized separation, C 18 Packing column inner diameter x length is 70 x 920mm, C 18 The particle size of the filler is 40-63 μm, eluted with methanol-water (76:24), the detection wavelength is 210nm, the required fraction is collected, the solvent is removed below 42°C, and further freeze-dried to obt...

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