CN101108872A - Plants natural base extractive and formulated product and use thereof - Google Patents
Plants natural base extractive and formulated product and use thereof Download PDFInfo
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Abstract
A botanical alkaloid extractive is provided, which is characterized in that the extractive is extracted from the and / or fruits, or solanum nigrum or solanum xanthocarpum. The invention also discloses a medicine of any form receivable pharmaceutically and mixedly produced by the botanical alkaloid extractive and the drug carriers receivable pharmaceutically. The extractive of the invention has scientific and reasonable extracting steps and has little impurity content and high content of the active ingredient solamargine with minimum of 90.0 per cent and maximum of 99.9 per cent. Certified by the test, the extractive preparation of the invention is safe and effective and controllable in quality.
Description
Technical field
The present invention relates to a kind of Chinese herbal medicine extract, particularly a kind of plant alkaloid extract the invention still further relates to the pharmaceutical preparation and the purposes of this extract.
Background technology
Black nightshade belongs to the Solanaceae Solanum, one to several years this plant of sward, cold, bitter, little sweet, have mild toxicity, return lung, stomach, urinary bladder channel, the effect of clearing heat and detoxicating, promoting blood circulation to remove blood stasis, inducing diuresis to remove edema, cough-relieving apophlegmatic is arranged, all there is distribution all parts of the country.In China 5 different shape types are arranged, yellow fruit black nightshade, purple stem black nightshade, green stem black nightshade, green arteries and veins spend black nightshade, brown arteries and veins to spend black nightshade less less, and main research is the wild black nightshade of northern China (SolanumnigrumL.) at present, belongs to purple stem black nightshade, but the black nightshade all herbal medicine.
The black nightshade herb contains the glycoside tetrahydroisoquinoline alkaloid, black nightshade polysaccharide, mineral substance, VITAMIN, pigment, amino acid etc., also contain ester, carboxylic compound, sterol, phenoloid (its main component is yellow unsaturated ester) in the extract of black nightshade berry, contain saponin(e and sapogenin in addition, and the chemistry that plays a crucial role is mainly alkaloid, is polysaccharide secondly.Report black nightshade total alkaloidss such as Liu Ying comprise solasonine, solamargine and B-solamargine, and Li Xiuxia etc. think that containing solamargine, solasonine, solancarpine and eggplant reaches alkali.
Chinese scholars studies confirm that Herba Solani Nigri extract has antitumor action, antibacterial and antivirus action, neuropharmacology effect, cardiovascular pharmacological effect, the effect of Digestive tract pharmacology, urinary system pharmacological action, genetic toxicity effect, internal secretion pharmacological action, the effect of antipyretic-antalgic pharmacology and some other pharmacological action.Its equal plant Australian eggplant (Solanum abiculare parst), Fruit of yellowfruit nightshade (Solanum xanthocar pumSchrad, et wendl) all contain similar composition, have similar pharmacological action.Now, which kind of extraction process can obtain the safe and effective dosage of effective constituent and preparation, then become the another emphasis of research by.
Summary of the invention
Technical problem to be solved by this invention is at the deficiencies in the prior art, and the plant alkaloid that a kind of foreign matter content is few, solamargine content is high extract is provided.
Another kind of technical problem to be solved by this invention provides the pharmaceutical preparation of said extracted thing.The present invention also provides the purposes of said extracted thing itself and pharmaceutical preparation.
Technical problem to be solved by this invention is to realize by following technical scheme.The present invention is a kind of plant alkaloid extract, is characterized in, it is from black nightshade herb and fruit, and perhaps Fruit of yellowfruit nightshade perhaps extracts in the Australian eggplant, and its extracting method step is as follows:
(1) gets black nightshade herb and fruit, perhaps Fruit of yellowfruit nightshade, perhaps Australian eggplant adds the 50-90% ethanolic soln and extracts 2-4 time, and each 4-8 doubly measures, reclaim ethanol, be concentrated into relative density 1.10～1.25, add 0.01～0.1mol/l acidic solution of 1～4 times of amount in the concentrated solution, fully stir, refrigeration was filtered more than 12 hours; Filter residue washs 1～3 time with 0.01～0.1mol/l acidic solution, and merging filtrate and washings are with low-pole or middle polarity resin absorption;
Perhaps,
Get black nightshade herb and fruit, perhaps Fruit of yellowfruit nightshade, perhaps Australian eggplant, with 0.01～0.1mol/l acidic solution dipping 2～4 times, 4～8 times of each consumptions, merge steeping fluid, with low-pole or middle polarity resin absorption at each 6～24 hours time;
(2) first water washes resin column/bed to effluent liquid neutrality after the resin absorption, colourless with alkaline ethanol solution flushing resin column/bed again to effluent liquid, perhaps colourless with basic solution flushing resin column/bed earlier, colourless to contain alcohol amount 5%～40% ethanol liquid flushing resin column/bed again to effluent liquid to effluent liquid; Discard washing fluid, carry out wash-out with 50-80% ethanol again, show colourless, collect elutriant, reclaim ethanol, transfer pH value to 8-10, leave standstill refrigeration, collecting precipitation with basic solution to there not being the alcohol flavor to elutriant;
(3) get the precipitation drying after, with 20～25 times of amount organic solvent dissolutions, add the chromatographic silica gel of 0.5～4 times of amount of precipitation capacity, mix thoroughly, evaporate to dryness is got the silica gel of mixing behind the sample, column chromatography for separation, chromatography column is adorned post, organic solvent wash-out with the organic solvent wet method, elutriant is collected in gradation, thin-layer chromatography detects, and merges the elutriant of thin-layer chromatography detection and solamargine sample point position consistency, reclaims organic solvent, evaporate to dryness, promptly;
Perhaps,
(3) get the precipitation drying after, add the bigger organic solvent of polarity of 5～20 times of amounts, stirred 10～30 minutes, leave standstill, filter, filtrate adds the low polar organic solvent of 2～6 times of amounts, stir, refining, filter, reclaim organic solvent, add the recrystallization organic solvent, carry out recrystallization, collect crystallization, promptly.
Technical problem to be solved by this invention can also further realize by following technical scheme.Above-described a kind of plant alkaloid extract is characterized in that the acidic solution described in the step (1) is selected from acetic acid, hydrochloric acid, sulfuric acid or citric acid solution; Basic solution described in the step (2) is selected from sodium hydroxide, calcium hydroxide, potassium hydroxide, magnesium hydroxide or the ammonia soln that concentration is 0.05-1.5mol/l, described alkaline ethanol solution is to contain the alkaline ethanol solution that is selected from sodium hydroxide, calcium hydroxide, potassium hydroxide, magnesium hydroxide or ammoniacal liquor that concentration is 0.05-0.15mol/l, and determining alcohol is 5%～40%.
Technical problem to be solved by this invention can also further realize by following technical scheme.Above-described a kind of plant alkaloid extract is characterized in that the dissolving described in the step (3) is selected from methyl alcohol, dehydrated alcohol, acetone, Virahol or propylene glycol with organic solvent; Described column chromatography wash-out is selected from the mixed phase that in methyl alcohol, dehydrated alcohol, acetone, propyl carbinol, chloroform, the ethyl acetate or 2-3 item form with arbitrary proportion with organic solvent; The organic solvent that described polarity is bigger is selected from methyl alcohol, ethanol, propyl carbinol, acetone or two mixed phase wherein; Described low polar organic solvent is selected from chloroform, methylene dichloride, ethylene dichloride, ethyl acetate, toluene or sherwood oil; Described recrystallization is selected from 40%～80% methyl alcohol, 40%～70% ethanol, perhaps 20%～60% acetone with organic solvent.
Technical problem to be solved by this invention can also further realize by following technical scheme.The invention discloses a kind of plant alkaloid extract formulation, be characterized in, it be the plant alkaloid extract and pharmaceutically the acceptable pharmaceutical carrier be mixed and made into the medicament of acceptable any formulation on the pharmaceutics.
Above-mentioned pharmaceutically acceptable carrier is meant the pharmaceutical carrier of pharmaceutical field routine, for example: thinner, vehicle and water etc., weighting agent such as starch, sucrose, lactose, Microcrystalline Cellulose etc.; Tackiness agent such as derivatived cellulose, alginate, gelatin and polyvinylpyrrolidone; Wetting agent such as glycerine; Disintegrating agent such as sodium starch glycolate, hydroxypropylcellulose, cross-linked carboxymethyl cellulose, agar, lime carbonate and sodium bicarbonate; Absorption enhancer such as quaternary ammonium compound; Tensio-active agent such as cetyl alcohol, sodium lauryl sulphate; Absorption carrier such as kaolin and soap clay; Lubricant such as talcum powder, calcium stearate and magnesium, micropowder silica gel and polyoxyethylene glycol etc.; Can also in composition, add other assistant agent such as flavouring agent, sweeting agent etc. in addition.
Preparation of the present invention can be applied to the patient who needs this treatment by the mode of oral, rectum or administered parenterally.Described oral, rectum and parenteral admin preparation are the described all oral or external preparations of pharmaceuticies such as tablet, capsule, sprays, pulvis, granule, oral liquid, dripping pill, pill, powder, suppository, sustained release preparation, controlled release preparation, suppository, gelifying agent, patch, cataplasma, film, syrup, elixir; Described injection formulations is injection, infusion solution, freeze dried injection, lipidosome injection, the described whole injection formulationss of pharmaceutics such as water or oiliness suspension agent, target administration injection, injectable emulsion; Preferred form is tablet, coated tablet, capsule, granule, oral liquid and injection.Various formulations can be according to the conventional production method preparation of pharmaceutical field.Extract is mixed with one or more carriers, be made into required formulation then.
Technical problem to be solved by this invention can also further realize by following technical scheme.Above-described a kind of plant alkaloid extract formulation is characterized in, in extract and pharmaceutical carrier mixing raw material, the weight percent content of extract is 0.1%-99.5%; Preferred 0.5%-95%.
Technical problem to be solved by this invention can also further realize by following technical scheme.Above-described a kind of plant alkaloid extract formulation is characterized in, described medicament is tablet or the capsule that extract adds weighting agent, disintegrating agent assembly; Or extract adds the slow releasing tablet or the capsule of weighting agent and hypromellose K4M assembly; Or extract is scattered in and obtains soft capsule in the oil phase; Or extract adds the injection of solubilizing agent or solubility promoter formation; Or extract is scattered in the injectable emulsion that obtains in the oil phase; Or extract adding pharmaceutical excipient is made suspension type injection liquid or freeze-drying injection powder pin.
Technical problem to be solved by this invention can also further realize by following technical scheme.Above-described a kind of plant alkaloid extract formulation is characterized in that described weighting agent is selected from lactose, Microcrystalline Cellulose, dextrin, starch or calcium phosphate; Disintegrating agent is selected from hydroxypropylcellulose, sodium starch glycolate, polyvinylpolypyrrolidone or croscarmellose sodium; Also optional tackiness agent, lubricant or the wetting agent of adding when film-making agent and capsule; Described oil phase is selected from soybean oil, poly(oxyethylene glycol) 400, Oleum Gossypii semen, peanut oil, sesame oil, Semen Maydis oil or sweet oil; When the system soft capsule, also can add solubilizing agent or latent solvent and oxidation inhibitor etc.; The used solubilizing agent of injection is selected from Soxylat A 25-7 Viscotrol C, tween or pluronic F-68; Solubility promoter can be selected dilute hydrochloric acid, ornithine hydrochloride, arginine monohydrochloride, Citric Acid, acidic amino acid or other acid organism and bisalt thereof for use, and perhaps alcohols comprises ethanol, propylene glycol and N.F,USP MANNITOL.
Technical problem to be solved by this invention can also further realize by following technical scheme.Above-described a kind of plant alkaloid extract formulation, be characterized in that the preparation method of described suspension type injection liquid is, with extract and Polysorbate 80 mix grind after, be dissolved into the aqueous solution of phosphoric acid potassium dihydrogen, dipotassium hydrogen phosphate, nipagin esters and Xylo-Mucine, make through grinding.
A kind of plant alkaloid extract of the present invention and preparation thereof can be used for preparing the medicine for the treatment of liver cancer, lung cancer and other tumours, and the medicine of preparation treatment inflammation.
Major ingredient in the plant alkaloid extract of the present invention is a solamargine, therefore hereinafter it is abbreviated as the solamargine extract.
Solamargine (Solamargine), its structural formula is as follows:
Preparation amount of application of the present invention can be according to variations such as the type of route of administration, patient age, body weight, body surface area, the disease of being treated and severity, and its safe and effective per daily dose can be:
The safe and effective dosage of people's clinical oral administration, rectum and parenteral admin: solamargine extract 0.1～10mg/kg body weight/day; The safe and effective dosage of people's clinical injection: solamargine extract 0.01～2mg/kg body weight/day; Can use by one or many.
Compared with prior art, the extraction step of extract of the present invention is scientific and reasonable, extract obtained in foreign matter content few, the content height of active ingredient solamargine wherein, the content of solamargine is minimum to reach 90.0%, reaches as high as 99.9%; The experiment proved that extract formulation of the present invention is safe and effective, and is quality controllable.
Embodiment
The following examples can make those skilled in the art more fully understand the present invention, but do not limit the present invention in any way.
Embodiment 1.A kind of plant alkaloid extract, it extracts from the black nightshade herb, and its extracting method step is as follows:
(1) get the black nightshade herb, add 50% ethanolic soln and extract 2 times, each 4 times of amounts reclaim ethanol, are concentrated into relative density 1.10, add the 0.01mol/l acidic solution of 1 times of amount in the concentrated solution, fully stir, and refrigeration was filtered more than 12 hours; Filter residue washs 1 time with the 0.01mol/l acidic solution, and merging filtrate and washings are with low-pole or middle polarity resin absorption;
(2) first water washes resin column/bed to effluent liquid neutrality after the resin absorption, colourless with alkaline ethanol solution flushing resin column/bed again to effluent liquid, perhaps colourless with basic solution flushing resin column/bed earlier, colourless to contain alcohol amount 5% ethanol liquid flushing resin column/bed again to effluent liquid to effluent liquid; Discard washing fluid, carry out wash-out with 50% ethanol again, show colourless, collect elutriant, reclaim ethanol, transfer pH value to 8, leave standstill refrigeration, collecting precipitation with basic solution to there not being the alcohol flavor to elutriant;
(3) get the precipitation drying after, with 20 times of amount organic solvent dissolutions, add the chromatographic silica gel of 0.5 times of amount of precipitation capacity, mix thoroughly, evaporate to dryness is got the silica gel of mixing behind the sample, column chromatography for separation, chromatography column is adorned post, organic solvent wash-out with the organic solvent wet method, elutriant is collected in gradation, thin-layer chromatography detects, and merges the elutriant of thin-layer chromatography detection and solamargine sample point position consistency, reclaims organic solvent, evaporate to dryness, promptly.
Embodiment 2.A kind of plant alkaloid extract, it extracts from Solanum nigrum fruit, and its extracting method step is as follows:
(1) get black nightshade herb and fruit, perhaps Fruit of yellowfruit nightshade, perhaps Australian eggplant adds 90% ethanolic soln and extracts 4 times, each 8 times of amounts reclaim ethanol, are concentrated into relative density 1.25, add the 0.1mol/l acidic solution of 4 times of amounts in the concentrated solution, fully stir, refrigeration was filtered more than 12 hours; Filter residue washs 3 times with the 0.1mol/l acidic solution, and merging filtrate and washings are with low-pole or middle polarity resin absorption;
(2) first water washes resin column/bed to effluent liquid neutrality after the resin absorption, colourless with alkaline ethanol solution flushing resin column/bed again to effluent liquid, perhaps colourless with basic solution flushing resin column/bed earlier, colourless to contain alcohol amount 40% ethanol liquid flushing resin column/bed again to effluent liquid to effluent liquid; Discard washing fluid, carry out wash-out with 80% ethanol again, show colourless, collect elutriant, reclaim ethanol, transfer pH value to 10, leave standstill refrigeration, collecting precipitation with basic solution to there not being the alcohol flavor to elutriant;
(3) get the precipitation drying after, with 25 times of amount organic solvent dissolutions, add the chromatographic silica gel of 4 times of amounts of precipitation capacity, mix thoroughly, evaporate to dryness is got the silica gel of mixing behind the sample, column chromatography for separation, chromatography column is adorned post, organic solvent wash-out with the organic solvent wet method, elutriant is collected in gradation, thin-layer chromatography detects, and merges the elutriant of thin-layer chromatography detection and solamargine sample point position consistency, reclaims organic solvent, evaporate to dryness, promptly.
Embodiment 3.A kind of plant alkaloid extract, it extracts from Fruit of yellowfruit nightshade, and its extracting method step is as follows:
(1) get Fruit of yellowfruit nightshade, add 70% ethanolic soln and extract 3 times, each 6 times of amounts reclaim ethanol, are concentrated into relative density 1.20, add the 0.05mol/l acidic solution of 2 times of amounts in the concentrated solution, fully stir, and refrigeration was filtered more than 12 hours; Filter residue washs 2 times with the 0.05mol/l acidic solution, and merging filtrate and washings are with low-pole or middle polarity resin absorption;
(2) first water washes resin column/bed to effluent liquid neutrality after the resin absorption, colourless with alkaline ethanol solution flushing resin column/bed again to effluent liquid, perhaps colourless with basic solution flushing resin column/bed earlier, colourless to contain alcohol amount 20% ethanol liquid flushing resin column/bed again to effluent liquid to effluent liquid; Discard washing fluid, carry out wash-out with 65% ethanol again, show colourless, collect elutriant, reclaim ethanol, transfer pH value to 9, leave standstill refrigeration, collecting precipitation with basic solution to there not being the alcohol flavor to elutriant;
(3) get the precipitation drying after, with 22 times of amount organic solvent dissolutions, add the chromatographic silica gel of 2 times of amounts of precipitation capacity, mix thoroughly, evaporate to dryness is got the silica gel of mixing behind the sample, column chromatography for separation, chromatography column is adorned post, organic solvent wash-out with the organic solvent wet method, elutriant is collected in gradation, thin-layer chromatography detects, and merges the elutriant of thin-layer chromatography detection and solamargine sample point position consistency, reclaims organic solvent, evaporate to dryness, promptly.
Embodiment 4.A kind of plant alkaloid extract, it extracts from Australian eggplant, and its extracting method step is as follows:
(1) get Australian eggplant, add 60% ethanolic soln and extract 3 times, each 5 times of amounts reclaim ethanol, are concentrated into relative density 1.15, add the 0.08mol/l acidic solution of 3 times of amounts in the concentrated solution, fully stir, and refrigeration was filtered more than 12 hours; Filter residue washs 2 times with the 0.08mol/l acidic solution, and merging filtrate and washings are with low-pole or middle polarity resin absorption;
(2) first water washes resin column/bed to effluent liquid neutrality after the resin absorption, colourless with alkaline ethanol solution flushing resin column/bed again to effluent liquid, perhaps colourless with basic solution flushing resin column/bed earlier, colourless to contain alcohol amount 10% ethanol liquid flushing resin column/bed again to effluent liquid to effluent liquid; Discard washing fluid, carry out wash-out with 60% ethanol again, show colourless, collect elutriant, reclaim ethanol, transfer pH value to 8.5, leave standstill refrigeration, collecting precipitation with basic solution to there not being the alcohol flavor to elutriant;
(3) get the precipitation drying after, add the bigger organic solvent of polarity of 5 times of amounts, stirred 10 minutes, leave standstill filtration, filtrate adds the low polar organic solvent of 2 times of amounts, stirs, and is refining, filters, and reclaims organic solvent, add the recrystallization organic solvent, carry out recrystallization, collect crystallization, promptly.
Embodiment 5.A kind of plant alkaloid extract, it extracts from black nightshade herb and fruit, and its extracting method step is as follows:
(1) get black nightshade herb and fruit, add 80% ethanolic soln and extract 2 times, each 7 times of amounts reclaim ethanol, are concentrated into relative density 1.22, add the 0.02mol/l acidic solution of 2.5 times of amounts in the concentrated solution, fully stir, and refrigeration was filtered more than 12 hours; Filter residue washs 3 times with the 0.02mol/l acidic solution, and merging filtrate and washings are with low-pole or middle polarity resin absorption;
(2) first water washes resin column/bed to effluent liquid neutrality after the resin absorption, colourless with alkaline ethanol solution flushing resin column/bed again to effluent liquid, perhaps colourless with basic solution flushing resin column/bed earlier, colourless to contain alcohol amount 30% ethanol liquid flushing resin column/bed again to effluent liquid to effluent liquid; Discard washing fluid, carry out wash-out with 70% ethanol again, show colourless, collect elutriant, reclaim ethanol, transfer pH value to 9.5, leave standstill refrigeration, collecting precipitation with basic solution to there not being the alcohol flavor to elutriant;
(3) get the precipitation drying after, add the bigger organic solvent of polarity of 20 times of amounts, stirred 30 minutes, leave standstill filtration, filtrate adds the low polar organic solvent of 6 times of amounts, stirs, and is refining, filters, and reclaims organic solvent, add the recrystallization organic solvent, carry out recrystallization, collect crystallization, promptly.
Embodiment 6.A kind of plant alkaloid extract, it extracts from black nightshade herb and fruit, and its extracting method step is as follows:
(1) get black nightshade herb and fruit, flood 2 times with the 0.01mol/l acidic solution, 4 times of each consumptions, merge steeping fluid, with low-pole or middle polarity resin absorption at each 6 hours time;
(2) first water washes resin column/bed to effluent liquid neutrality after the resin absorption, colourless with alkaline ethanol solution flushing resin column/bed again to effluent liquid, perhaps colourless with basic solution flushing resin column/bed earlier, colourless to contain alcohol amount 25% ethanol liquid flushing resin column/bed again to effluent liquid to effluent liquid; Discard washing fluid, carry out wash-out with 75% ethanol again, show colourless, collect elutriant, reclaim ethanol, transfer pH value to 9, leave standstill refrigeration, collecting precipitation with basic solution to there not being the alcohol flavor to elutriant;
(3) get the precipitation drying after, add the bigger organic solvent of polarity of 12 times of amounts, stirred 20 minutes, leave standstill filtration, filtrate adds the low polar organic solvent of 4 times of amounts, stirs, and is refining, filters, and reclaims organic solvent, add the recrystallization organic solvent, carry out recrystallization, collect crystallization, promptly.
Embodiment 7.A kind of plant alkaloid extract, it extracts from Fruit of yellowfruit nightshade, and its extracting method step is as follows:
(1) get Fruit of yellowfruit nightshade, flood 4 times with the 0.1mol/l acidic solution, 8 times of each consumptions, merge steeping fluid, with low-pole or middle polarity resin absorption at each 24 hours time;
(2) first water washes resin column/bed to effluent liquid neutrality after the resin absorption, colourless with alkaline ethanol solution flushing resin column/bed again to effluent liquid, perhaps colourless with basic solution flushing resin column/bed earlier, colourless to contain alcohol amount 35% ethanol liquid flushing resin column/bed again to effluent liquid to effluent liquid; Discard washing fluid, carry out wash-out with 55% ethanol again, show colourless, collect elutriant, reclaim ethanol, transfer pH value to 8, leave standstill refrigeration, collecting precipitation with basic solution to there not being the alcohol flavor to elutriant;
(3) get the precipitation drying after, add the bigger organic solvent of polarity of 15 times of amounts, stirred 25 minutes, leave standstill filtration, filtrate adds the low polar organic solvent of 3 times of amounts, stirs, and is refining, filters, and reclaims organic solvent, add the recrystallization organic solvent, carry out recrystallization, collect crystallization, promptly.
Embodiment 8.A kind of plant alkaloid extract, it extracts from Australian eggplant, and its extracting method step is as follows:
(1) get Australian eggplant, flood 3 times with the 0.05mol/l acidic solution, 6 times of each consumptions, merge steeping fluid, with low-pole or middle polarity resin absorption at each 12 hours time;
(2) first water washes resin column/bed to effluent liquid neutrality after the resin absorption, colourless with alkaline ethanol solution flushing resin column/bed again to effluent liquid, perhaps colourless with basic solution flushing resin column/bed earlier, colourless to contain alcohol amount 25% ethanol liquid flushing resin column/bed again to effluent liquid to effluent liquid; Discard washing fluid, carry out wash-out with 70% ethanol again, show colourless, collect elutriant, reclaim ethanol, transfer pH value to 9, leave standstill refrigeration, collecting precipitation with basic solution to there not being the alcohol flavor to elutriant;
(3) get the precipitation drying after, add the bigger organic solvent of polarity of 10 times of amounts, stirred 15 minutes, leave standstill filtration, filtrate adds the low polar organic solvent of 5 times of amounts, stirs, and is refining, filters, and reclaims organic solvent, add the recrystallization organic solvent, carry out recrystallization, collect crystallization, promptly.
Embodiment 9.A kind of plant alkaloid extract, it extracts from the black nightshade herb, and its extracting method step is as follows:
(1) get the black nightshade herb, flood 3 times with the 0.08mol/l acidic solution, 5 times of each consumptions, merge steeping fluid, with low-pole or middle polarity resin absorption at each 20 hours time;
(2) first water washes resin column/bed to effluent liquid neutrality after the resin absorption, colourless with alkaline ethanol solution flushing resin column/bed again to effluent liquid, perhaps colourless with basic solution flushing resin column/bed earlier, colourless to contain alcohol amount 30% ethanol liquid flushing resin column/bed again to effluent liquid to effluent liquid; Discard washing fluid, carry out wash-out with 75% ethanol again, show colourless, collect elutriant, reclaim ethanol, transfer pH value to 9, leave standstill refrigeration, collecting precipitation with basic solution to there not being the alcohol flavor to elutriant;
(3) get the precipitation drying after, with 22 times of amount organic solvent dissolutions, add the chromatographic silica gel of 3 times of amounts of precipitation capacity, mix thoroughly, evaporate to dryness is got the silica gel of mixing behind the sample, column chromatography for separation, chromatography column is adorned post, organic solvent wash-out with the organic solvent wet method, elutriant is collected in gradation, thin-layer chromatography detects, and merges the elutriant of thin-layer chromatography detection and solamargine sample point position consistency, reclaims organic solvent, evaporate to dryness, promptly.
Embodiment 10.In present embodiment 1, the content of solamargine is 90.0% in this extract; Acidic solution described in the step (1) is an acetum; Basic solution described in the step (2) is that concentration is the sodium hydroxide solution of 0.05mol/l, and described alkaline ethanol solution is to contain the alkaline ethanol solution that concentration is the sodium hydroxide of 0.05mol/l, and determining alcohol is 5%; Dissolving organic solvent described in the step (3) is a methyl alcohol; Described column chromatography wash-out organic solvent is in methyl alcohol, dehydrated alcohol, acetone, propyl carbinol, chloroform, the ethyl acetate.
Embodiment 11.In present embodiment 2, the content of solamargine is 95% in this extract; Acidic solution described in the step (1) is a hydrochloric acid soln; Basic solution described in the step (2) is that concentration is the aqua calcis of 1.5mol/l, and described alkaline ethanol solution is to contain the alkaline ethanol solution that concentration is the calcium hydroxide of 0.15mol/l, and determining alcohol is 40%; Dissolving organic solvent described in the step (3) is a dehydrated alcohol; Described column chromatography wash-out organic solvent is 2 mixed phase that form with arbitrary proportion in methyl alcohol, dehydrated alcohol, acetone, propyl carbinol, chloroform, the ethyl acetate.
Embodiment 12.In present embodiment 3, the content of solamargine is 99.9% in this extract; Acidic solution described in the step (1) is a sulphuric acid soln; Basic solution described in the step (2) is selected from the potassium hydroxide solution that concentration is 1.0mol/l, and described alkaline ethanol solution is to contain to be selected from the alkaline ethanol solution that concentration is the potassium hydroxide of 0.10mol/l, and determining alcohol is 20%; Dissolving organic solvent described in the step (3) is an acetone; Described column chromatography wash-out organic solvent is 3 mixed phase that form with arbitrary proportion in methyl alcohol, dehydrated alcohol, acetone, propyl carbinol, chloroform, the ethyl acetate.
Embodiment 13.In present embodiment 4, the content of solamargine is 91.0% in this extract; Acidic solution described in the step (1) is a citric acid solution; Basic solution described in the step (2) is selected from the magnesium hydroxide solution that concentration is 0.10mol/l, and described alkaline ethanol solution is to contain to be selected from the alkaline ethanol solution that concentration is the magnesium hydroxide of 0.08mol/l, and determining alcohol is 10%; The organic solvent that polarity described in the step (3) is bigger is methyl alcohol or ethanol or propyl carbinol or acetone; Described low polar organic solvent is chloroform or methylene dichloride; Described recrystallization organic solvent is 40%～80% methyl alcohol.
Embodiment 14.In present embodiment 5, the content of solamargine is 94.0% in this extract; Acidic solution described in the step (1) is a citric acid solution; Basic solution described in the step (2) is that concentration is the ammonia soln of 0.50mol/l, and described alkaline ethanol solution is to contain to be selected from the alkaline ethanol solution that concentration is the ammoniacal liquor of 0.12mol/l, and determining alcohol is 30%; The organic solvent that polarity described in the step (3) is bigger is methyl alcohol and alcoholic acid mixed phase; Described low polar organic solvent is ethylene dichloride or ethyl acetate; Described recrystallization organic solvent is 40%～70% ethanol.
Embodiment 15.In present embodiment 6, the content of solamargine is 98% in this extract; Acidic solution described in the step (1) is an acetum; Basic solution described in the step (2) is selected from the sodium hydroxide solution that concentration is 1.2mol/l, and described alkaline ethanol solution is to contain to be selected from the alkaline ethanol solution that concentration is the sodium hydroxide of 0.07mol/l, and determining alcohol is 35%; The organic solvent that polarity described in the step (3) is bigger is the mixed phase of propyl carbinol and acetone; Described low polar organic solvent is toluene or sherwood oil; Described recrystallization organic solvent is 20%～60% acetone.
Embodiment 16.In present embodiment 7, the content of solamargine is 97% in this extract; Acidic solution described in the step (1) is a hydrochloric acid soln; Basic solution described in the step (2) is that concentration is the aqua calcis of 0.80mol/l, and described alkaline ethanol solution is to contain to be selected from the alkaline ethanol solution that concentration is the calcium hydroxide of 0.08mol/l, and determining alcohol is 20%; The organic solvent that polarity described in the step (3) is bigger is the mixed phase of methyl alcohol and propyl carbinol; Described low polar organic solvent is a sherwood oil; Described recrystallization organic solvent is 60% methyl alcohol.
Embodiment 17.In present embodiment 8, the content of solamargine is 93.0% in this extract; Acidic solution described in the step (1) is a sulphuric acid soln; Basic solution described in the step (2) is selected from the potassium hydroxide solution that concentration is 1.1mol/l, and described alkaline ethanol solution is to contain to be selected from the alkaline ethanol solution that concentration is the potassium hydroxide of 0.14mol/l, and determining alcohol is 30%; The organic solvent that polarity described in the step (3) is bigger is the mixed phase of ethanol and acetone; Described low polar organic solvent is a chloroform; Described recrystallization organic solvent is 55% ethanol.
Embodiment 18.In present embodiment 9, the content of solamargine is 99.0% in this extract; Acidic solution described in the step (1) is a citric acid solution; Basic solution described in the step (2) is selected from the magnesium hydroxide solution that concentration is 1.3mol/l, and described alkaline ethanol solution is to contain to be selected from the alkaline ethanol solution that concentration is the magnesium hydroxide of 0.14mol/l, and determining alcohol is 28%; Dissolving organic solvent described in the step (3) is acetone or Virahol or propylene glycol; Described column chromatography wash-out organic solvent is in methyl alcohol, dehydrated alcohol, acetone, propyl carbinol, chloroform, the ethyl acetate.
Embodiment 19.A kind of as embodiment 1 described plant alkaloid extract formulation, it be the plant alkaloid extract and pharmaceutically the acceptable pharmaceutical carrier be mixed and made into oral preparations.
Embodiment 20.A kind ofly get described extract as embodiment 2 described plant alkaloid extract formulations, add weighting agent, tablet is made in the disintegrating agent assembly, in mixing raw material, the weight percent content of extract is 5%.
Embodiment 21.A kind ofly get described extract as embodiment 3 described plant alkaloid extract formulations, add weighting agent, capsule is made in the disintegrating agent assembly, in mixing raw material, the weight percent content of extract is 20%.
Embodiment 22.A kind ofly get described extract as embodiment 4 described plant alkaloid extract formulations, add weighting agent and hypromellose K4M assembly and make slow releasing tablet, in mixing raw material, the weight percent content of extract is 5%.
Embodiment 23.A kind ofly get described extract as embodiment 5 described plant alkaloid extract formulations, add weighting agent and hypromellose K4M assembly and make slow releasing capsule, in mixing raw material, the weight percent content of extract is 10%.
Embodiment 24.A kind ofly get described extract as embodiment 1 described plant alkaloid extract formulation, be scattered in and obtain soft capsule in the oil phase, in mixing raw material, the weight percent content of extract is 20%.
Embodiment 25.A kind ofly get described extract as embodiment 2 described plant alkaloid extract formulations, the injection that solubilizing agent or solubility promoter form, in mixing raw material, the weight percent content of extract is 0.1%.
Embodiment 26.A kind ofly get described extract as embodiment 3 described plant alkaloid extract formulations, be scattered in the injectable emulsion that obtains in the oil phase and add, in mixing raw material, the weight percent content of extract is 0.2%.
Embodiment 27.A kind ofly get described extract as embodiment 4 described plant alkaloid extract formulations, add pharmaceutical excipient and make the suspension type injection liquid, in mixing raw material, the weight percent content of extract is 0.5%.
Embodiment 28.A kind ofly get described extract as embodiment 5 described plant alkaloid extract formulations, add pharmaceutical excipient and make freeze-drying injection powder pin, in mixing raw material, the weight percent content of extract is 10%.
Embodiment 29.A kind of as embodiment 1 described plant alkaloid extract formulation, with extract and Polysorbate 80 mix grind after, be dissolved into the aqueous solution of phosphoric acid potassium dihydrogen, dipotassium hydrogen phosphate, nipagin esters and Xylo-Mucine, make the suspension type injection liquid through grinding, in mixing raw material, the weight percent content of extract is 0.3%.Be used for the treatment of liver cancer, lung cancer and other tumours and inflammation.
Embodiment 30.The agent of solamargine extract capsule.
Solamargine extract 10-60 gram
Microcrystalline Cellulose 30-50 gram
Lactose 30-50 gram
Sodium starch glycolate 5-10 gram
2%HPMCE5 solution (containing 5% tween 80) is an amount of
Magnesium Stearate 1-5 gram
Its preparation method is to get the solamargine extract and be dissolved in small amount of ethanol; Microcrystalline Cellulose, lactose, sodium starch glycolate are crossed 100 mesh sieves respectively and are mixed, and add solamargine extract ethanolic soln and fully stirring, and ethanol is all volatilized; HPMC solution with tween 80 is tackiness agent system softwood, crosses 20 mesh sieve system particles, and wet granular is in 50--60 ℃ of baking oven forced air drying; Dried particle is crossed the whole grain of 20 mesh sieves, with the Magnesium Stearate mixing, in can 1000 capsules.
Embodiment 31.Solamargine extract tablet.
Solamargine extract 10-60 gram
Microcrystalline Cellulose 30-50 gram
Lactose 30-50 gram
Sodium starch glycolate 5-10 gram
2%HPMCE5 solution (containing 5% tween 80) is an amount of
Magnesium Stearate 1-5 gram
Its preparation method is to get the solamargine extract and be dissolved in small amount of ethanol; Microcrystalline Cellulose, lactose, sodium starch glycolate are crossed 100 mesh sieves respectively and are mixed, and add solamargine extract ethanolic soln and fully stirring, and ethanol is all volatilized; HPMC solution with tween 80 is tackiness agent system softwood, crosses 20 mesh sieve system particles, and wet granular is in 50--60 ℃ of baking oven forced air drying; Dried particle is crossed the whole grain of 20 mesh sieves, with the Magnesium Stearate mixing, is pressed into 1000.
Embodiment 32.Solamargine extract slow-release capsule.
Solamargine extract 60mg
Microcrystalline Cellulose 20mg
Hypromellose K4M 80mg
3% HPMC (E5) aqueous solution is an amount of
Talcum powder 4mg
Solamargine extract, Microcrystalline Cellulose, hypromellose K4M are crossed 60 mesh sieves and be mixed, add 3% HPMC (E5) aqueous solution and make softwood in right amount, cross 20 mesh sieves and granulate.40-50 ℃ of baking oven forced air drying.Dried particle is crossed the whole grain of 20 mesh sieves, adds the talcum powder of recipe quantity, mixes.Press No. 1 capsule of recipe quantity can, every contains solamargine extract 60mg.
Embodiment 33.Solamargine extract slow-release tablet.
Solamargine extract 100mg
Lactose 20mg
Hypromellose K4M 80mg
3% HPMC (E5) aqueous solution is an amount of
Talcum powder 4mg
Solamargine extract, lactose, hypromellose K4M are crossed 60 mesh sieves and be mixed, add 3% HPMC (E5) aqueous solution and make softwood in right amount, cross 20 mesh sieves and granulate.40-50 ℃ of baking oven forced air drying.Dried particle is crossed the whole grain of 20 mesh sieves, adds the talcum powder of recipe quantity, mix, and compressing tablet, every contains solamargine extract 100mg.
Above-mentioned example also can be selected other auxiliary material for use, and disintegrating agent is as hydroxypropylated starch, hydroxypropylcellulose, sodium starch glycolate, calcium carboxymethylcellulose, polyvinylpolypyrrolidone, croscarmellose sodium etc.; Weighting agent is as lactose, sucrose, N.F,USP MANNITOL, Microcrystalline Cellulose, dextrin, starch, calcium phosphate, secondary calcium phosphate, calcium sulfate, lime carbonate, cyclodextrin, micro mist Mierocrystalline cellulose etc.; Wetting agent and tackiness agent are as pregelatinized Starch, polyvidone, Xylo-Mucine, hypromellose; Lubricant is as talcum powder, stearic acid, Magnesium Stearate, calcium stearate, micropowder silica gel, hydrogenated vegetable oil, Macrogol 4000 and 6000; Wetting agent is as sodium lauryl sulphate, tween 80; Framework material is as hypromellose, ethyl cellulose etc.
Embodiment 34.The solamargine extract soft capsule.
Every of content contains the glue shell
Solamargine extract 150mg gelatin 46.00%
Soybean oil 0.5ml glycerine 17.82%
Water 36.18%
Get the solamargine extract and be dissolved in the soybean oil, this solution is made soft capsule.Every contains solamargine extract 150mg.
Soft capsule of the present invention also can be selected following auxiliary material for use: solvent is as poly(oxyethylene glycol) 400, Oleum Gossypii semen, peanut oil, sesame oil, Semen Maydis oil, olive wet goods; Solubilizing agent or latent solvent are as tween 80, Soxylat A 25-7 Viscotrol C, peruscabin, ethyl lactate etc.; Oxidation inhibitor is as Tenox PG, t-butyl phenol (BHT), vitamin-E etc.The ratio of gelatin, glycerine and water can suitably be regulated in the glue shell, is advisable 1: 0.3～0.4: 0.7～1.4 as gelatin/glycerin/water three's ratio, also can add other compositions in the glue shell, as sanitas: P-hydroxybenzoic acid first, second, third, butyl ester etc.; Softening agent such as sorbyl alcohol etc.; Stablizer such as gum arabic etc.; Opalizer is as titanium dioxide, barium sulfate, precipitated calcium carbonate etc.
Embodiment 35.Solamargine extract injectable emulsion.
Solamargine extract 5g
Soybean oil 50g
Soybean phospholipid 12g
Glycerine 25g
Water for injection adds to 1000ml
Under nitrogen gas stream, fabaceous lecithin is added the soybean oil stirring make its dissolving, add glycerine and solamargine extract stirring and dissolving, slowly add water for injection under the agitation condition, through the high pressure dispersing emulsification machine emulsification of two steps; Still under nitrogen gas stream, with No. 4 sintered glass funnel filtration under diminished pressure, and lid is rolled in bottling under nitrogen gas stream, and elder generation,, cools off promptly towards hot water after sterilization finishes again in 121 ℃ of sterilizations 15 minutes gradually through preheating.
The following auxiliary material of also optional usefulness in this example: oil for injection is as ethyl oleate, poly(oxyethylene glycol) 400, Oleum Gossypii semen, peanut oil, sesame oil, Semen Maydis oil, sweet oil, Isopropyl myristate etc.; Oxidation inhibitor is as Tenox PG, t-butyl phenol (BHT), vitamin-E etc.; Tensio-active agent is as Tweens, Soxylat A 25-7 Viscotrol C, phospholipid, pluronic etc.
Embodiment 36.The solamargine extract injection.
Solamargine extract 20mg
Soxylat A 25-7 Viscotrol C 1.0mg
Dehydrated alcohol 5.0mg
Water for injection adds to 5.0mL
The solamargine extract is dissolved in dehydrated alcohol, add 20% Soxylat A 25-7 Viscotrol C (CremophorELP), mixing, reduction vaporization is removed ethanol, add an amount of water for injection and be mixed into clear solution, through 0.22 μ m filtering with microporous membrane, coating-dividing sealing, in 100 ℃ of flowing steam sterilizations 30 minutes promptly, every contains solamargine extract 20mg.
Embodiment 37.The solamargine extract injection.
Solamargine extract 20.0g
Arginine monohydrochloride 22.2g
Sodium-chlor is an amount of
Water for injection adds to 2000mL
Get solamargine extract and arginine monohydrochloride and put in the appropriate vessel, add injection water 1800ml, stir, ultrasonic to dissolving, add the sodium-chlor stirring and make dissolving; , add water for injection to 2000ml, through 0.22 μ m filtering with microporous membrane, coating-dividing sealing, in 100 ℃ of flowing steam sterilizations 30 minutes promptly, every contains solamargine extract 20mg.
Embodiment 38.Injection solamargine extract powder injection.
Solamargine extract 20.0g
Ornithine hydrochloride 22.2g
N.F,USP MANNITOL 32.0g
Water for injection is to 2000ml
Get solamargine extract and arginine and put in the appropriate vessel, add injection water 1800ml, stir, ultrasonic to dissolving, add the N.F,USP MANNITOL stirring and make dissolving; Add needle-use activated carbon by 0.1%, stirred 30 minutes, and took off the charcoal suction filtration in the container of cleaning, add water for injection to 2000ml through the titanium core, solution stirring was made evenly in 5 minutes, again through 0.22 μ m filtering with microporous membrane, the filtrate can in the 7ml cillin bottle, every bottle of 2ml, butyl rubber bung beyond the Great Wall partly then, deliver on the flaggy in the freeze drying box, insert temp probe, close chamber door.Press the freeze-drying curve lyophilize, the final drying temperature is more than 35 ℃ and kept 2 hours.Close plug, venting, outlet rolls lid.
Embodiment 39.Solamargine extract suspendible type injection.
Solamargine extract 20mg
Xylo-Mucine 10mg
Polysorbate 80 0.1mg
Ethyl p-hydroxybenzoate 0.5mg
Propylben 0.5mg
Potassium primary phosphate 16.7mg
Dipotassium hydrogen phosphate 1.7mg
Water for injection adds to 2ml
The solamargine extract is carried out comminution by gas stream, get the following micro mist of particle diameter 10 μ m.Potassium primary phosphate and dipotassium hydrogen phosphate are dissolved in the water for injection, add ethyl p-hydroxybenzoate and propyl ester, add Xylo-Mucine again, make whole dissolvings under 60 ℃ of conditions.Solamargine extract after the micronization is placed container, add Polysorbate 80 and be ground into thin pasty state, above-mentioned solution is added gradually, after stirring, grind 5 to 10 times through colloidal mill.Routinely measuring method measure content qualified after, be divided in the ampoule, in 100 ℃ of flowing steam sterilizations 30 minutes promptly, every contains solamargine extract 20mg.
Injection of the present invention also can be selected following auxiliary material for use: solubilizing agent is as Tweens, pluronic F-68, Soxylat A 25-7 castor-oil plant wet goods; Solubility promoter is as amides such as urea, ethanamide, thiocarbamide, benzamide etc., the compound of hydroxyl or carboxyl such as sucrose, Citric Acid and sodium salt thereof, lactic acid, sodium salicylate etc.; Suspending agent is as Xylo-Mucine, polyvidone, HPMC etc.; Sanitas is as: Metagin, second, third and butyl ester; The pH regulator agent is as Citric Acid and citrate, phosphoric acid salt etc.; Solvent is as water for injection, injection ethanol, propylene glycol etc.
Embodiment 40.The test of the solamargine extract oral pharmacological toxicology research that makes by embodiment 3.
(1) pharmacodynamic study
1, solamargine is to the influence of S180 sarcoma transplanted tumor tumour inhibiting rate
1.1 experimental technique
Aseptic condition extracts 8 days ascites cells suspension of growth in the KM mouse peritoneal down, and cancer cells suspension and sterile saline are got 50 of KM mouse by dilution in 1: 3, and the cancer cells suspension 0.2ml after the inoculation dilution is subcutaneous in armpit.Be divided into 7 groups behind the 24h at random, be that model group, 5-Fu organize (25mg/kg) and 1,2,3,4,5 groups of solamargines (0.6mg/kg, 1.2mg/kg, 2.4mg/kg, 4.8mg/kg, 9.6mg/kg), every group 10, irritate stomach respectively and give relative medicine (model group is irritated stomach and given isometric(al) distilled water), put to death mouse after 8 days, strip tumour, weigh, calculate tumour inhibiting rate by following formula.
1.2 experimental result
Mouse inoculation S
180After the tumour, 3 days subcutaneous can contact tumour, dissects exemplary embodiment lock after 8 days all greater than 1g, and 5 dosage groups of solamargine mouse tumor soaks into scope all less than model group, and the knurl body is easily peeled off, and tumor weight obviously alleviates (P＜0.05, P＜0.01).The results are shown in Table 1.
Table 1 solamargine is to S
180The influence of transplanted solid tumor tumour inhibiting rate (
N=10)
| Group | Dosage (mg/kg) | Knurl heavy (g) | Tumour inhibiting rate (%) |
| 4 groups of 3 groups of solamargines of model group 5-Fu group 2 groups of solamargines of 1 group of solamargine of solamargine | - 25 0.6 1.2 2.4 4.8 | 1.26±0.42 0.41±0.31 ** 0.81±0.49 * 0.73±0.37 ** 0.61±0.41 ** 0.56±0.42 ** | - 67.5 35.7 42.1 51.6 55.6 |
| 5 groups of solamargines | 9.6 | 0.50±0.34 ** | 60.3 |
Annotate: compare with model group,
*P＜0.01.
2, solamargine is to the influence of Hep liver cancer transplanted tumor tumour inhibiting rate
2.1 experimental technique
Aseptic condition extracts 8 days ascites cells suspension of growth in the KM mouse peritoneal down, and cancer cells suspension and sterile saline are got 50 of KM mouse by dilution in 1: 3, and the cancer cells suspension 0.2ml after the inoculation dilution is subcutaneous in armpit.Be divided into 5 groups behind the 24h at random, be that model group, 5-Fu organize (25mg/kg) and the basic, normal, high dosage group of solamargine (1.2mg/kg, 2.4mg/kg, 4.8mg/kg), irritate stomach respectively and give relative medicine (model group is irritated stomach and given isometric(al) distilled water), put to death mouse after 8 days, strip tumour, weigh, calculate tumour inhibiting rate by following formula.
2.2 experimental result
After the mouse inoculation Hep liver cancer tumour, subcutaneously in the time of four days can contact tumour, dissect exemplary embodiment lock after 8 days all greater than 1g, three dosage groups of solamargine mouse tumor soaks into scope all less than model group, the knurl body is easily peeled off, and tumor weight obviously alleviates (P＜0.05,0.01).Tumour inhibiting rate is all greater than 30%.The results are shown in Table 2.
Table 2 solamargine to the influence of Hep transplanted solid tumor tumour inhibiting rate (
)
| Group | Dosage (mg/kg) | Number of animals (n) | Knurl heavy (g) | Tumour inhibiting rate (%) |
| Dosage group solamargine high dose group in the model group 5-Fu group solamargine low dose group solamargine | - 25 1.2 2.4 4.8 | 10 10 10 10 10 | 1.40±0.56 0.51±0.36 ** 0.75±0.41 ** 0.62±0.39 ** 0.54±0.41 ** | - 63.6 46.4 55.7 61.4 |
Annotate: compare with model group,
*P＜0.05,
*P＜0.01.
3, to the influence of mice auricle swelling
3.1 experimental technique
Get 50 of normal ICR mouse, body weight 25～30g, male.Be divided into 5 groups at random, every group of 10 mouse, promptly blank group, acetylsalicylic acid group: (110mg/kg) with the basic, normal, high dosage group of solamargine (1.2mg/kg, 2.4mg/kg, 4.8mg/kg), irritate stomach respectively and give relative medicine (blank group is irritated stomach and given isometric(al) distilled water), once a day, continuous 3 days.Behind the administration 40min on the 3rd, be applied to two sides, ear front and back, a mouse left side with 2% Oleum Tiglii 0.05ml, after causing scorching 4h, put to death mouse, two ears about cutting along the auricle baseline, (diameter 9mm) takes off round auricle at same position respectively with punch tool, scales/electronic balance weighing as auricular concha swelling degree, and calculates the swelling percentage with the difference of mouse left and right sides auricular concha weight.
3.2 experimental result
Each dosage group of solamargine all can alleviate Oleum Tiglii and cause the mice ear degree, reduces the swelling rate, has compared significant difference (p＜0.05, p＜0.01) with the blank group.Show that solamargine has anti-inflammatory action, the results are shown in Table 3.
Table 3 solamargine to the influence of Oleum Tiglii induced mice ear swelling (
)
| Group | Dosage (mg/kg) | Number of animals (only) | Auricular concha swelling degree (mg) | The swelling rate |
| Dosage group solamargine high dose group in the blank group acetylsalicylic acid group solamargine low dose group solamargine | - 110 1.2 2.4 4.8 | 10 10 10 10 10 | 20.71±8.52 11.24±6.14 * 17.53±4.29 15.34±6.28 12.38±5.18 * | 123.2±42.2 65.8±32.5 ** 106.7±26.4 90.9±35.6 76.5±31.9 * |
Annotate: compare with the blank group,
*P＜0.05,
*P＜0.01.
In sum, the solamargine gastric infusion is to S
180, Hep liver cancer transplanted tumor has stronger antitumor action, solamargine also has certain anti-inflammatory action simultaneously.
(2) toxicological study
Acute toxicity test
Through acute toxicity test: gastric infusion of solamargine, its LD
1Dosage is 0.1g/kg, and being folded to the people is 10mg/kg.
Embodiment 41.The test of the solamargine extract injection formulations pharmacological toxicology research that makes by embodiment 3.
(1) pharmacodynamic study
1, the injection solamargine is to S
180The influence of sarcoma transplanted tumor tumour inhibiting rate
1.1 experimental technique
Aseptic condition extracts 8 days ascites cells suspension of growth in the KM mouse peritoneal down, and cancer cells suspension and sterile saline are got 50 of KM mouse by dilution in 1: 3, and the cancer cells suspension 0.2ml after the inoculation dilution is subcutaneous in armpit.Be divided into 7 groups behind the 24h at random, be that model group, 5-Fu organize (25mg/kg) and 1,2,3,4,5 groups of solamargines (0.03mg/kg, 0.06mg/kg, 0.12mg/kg, 0.24mg/kg, 0.48mg/kg), every group 10, intravenous injection gives relative medicine (the model group injection gives isometric(al) physiological saline) respectively, put to death mouse after 8 days, strip tumour, weigh, calculate tumour inhibiting rate by following formula.
1.2 experimental result
Mouse inoculation S
180After the tumour, 3 days subcutaneous can contact tumour, dissects exemplary embodiment lock after 8 days all greater than 1g, and 5 dosage groups of solamargine mouse tumor soaks into scope all less than model group, and the knurl body is easily peeled off, and tumor weight obviously alleviates (P＜0.01).The results are shown in Table 1.
Table 1 solamargine to the influence of S180 transplanted solid tumor tumour inhibiting rate (
N=10)
| Group | Dosage (mg/kg) | Knurl heavy (g) | Tumour inhibiting rate (%) |
| 5 groups of 4 groups of solamargines of model group 5-Fu group 3 groups of solamargines of 2 groups of solamargines of 1 group of solamargine of solamargine | - 25 0.03 0.06 0.12 0.24 0.48 | 1.44±0.55 0.56±0.30 ** 0.87±0.41 * 0.71±0.49 ** 0.67±0.35 ** 0.64±0.30 ** 0.57±0.38 ** | - 61.1 39.6 50.7 53.5 55.6 60.4 |
Annotate: compare with model group,
*P＜0.05,
*P＜0.01.
2, the injection solamargine is to the influence of Hep liver cancer transplanted tumor tumour inhibiting rate
2.1 experimental technique
Aseptic condition extracts 8 days ascites cells suspension of growth in the KM mouse peritoneal down, and cancer cells suspension and sterile saline are got 50 of KM mouse by dilution in 1: 3, and the cancer cells suspension 0.2ml after the inoculation dilution is subcutaneous in armpit.Be divided into 5 groups behind the 24h at random, be that model group, 5-Fu organize (25mg/kg) and the basic, normal, high dosage group of solamargine (0.06mg/kg, 0.12mg/kg, 0.24mg/kg), intravenous injection gives relative medicine (the model group injection gives isometric(al) physiological saline) respectively, put to death mouse after 8 days, strip tumour, weigh, calculate tumour inhibiting rate by following formula.
2.2 experimental result
After the mouse inoculation Hep liver cancer tumour, subcutaneously in the time of four days can contact tumour, dissect exemplary embodiment lock after 8 days all greater than 1g, three dosage groups of solamargine mouse tumor soaks into scope all less than model group, the knurl body is easily peeled off, and tumor weight obviously alleviates (P＜0.05,0.01).Tumour inhibiting rate is all greater than 30%.The results are shown in Table 2.
Table 2 solamargine to the influence of Hep transplanted solid tumor tumour inhibiting rate (
)
| Group | Dosage (mg/kg) | Number of animals (n) | Knurl heavy (g) | Tumour inhibiting rate (%) |
| Dosage group solamargine high dose group in the model group 5-Fu group solamargine low dose group solamargine | - 25 0.06 0.12 0.24 | 10 10 10 10 10 | 1.65±0.42 0.55±0.23 ** 0.81±0.34 ** 0.770.20 ** 0.65±0.33 ** | - 66.7 50.9 53.3 60.6 |
Annotate: compare with model group,
*P＜0.05,
*P＜0.01.
3, to the influence of mice auricle swelling
3.1 experimental technique
Get 50 of normal ICR mouse, body weight 25～30g, male.Be divided into 5 groups at random, every group of 10 mouse, promptly blank group, acetylsalicylic acid group: (110mg/kg) with the basic, normal, high dosage group of solamargine (0.06mg/kg, 0.12mg/kg, 0.24mg/kg), intravenous injection gives relative medicine (blank group injection gives isometric(al) physiological saline) respectively, once a day, continuous 3 days.Behind the administration 40min on the 3rd, be applied to two sides, ear front and back, a mouse left side with 2% Oleum Tiglii 0.05ml, after causing scorching 4h, put to death mouse, two ears about cutting along the auricle baseline, (diameter 9mm) takes off round auricle at same position respectively with punch tool, scales/electronic balance weighing as auricular concha swelling degree, and calculates the swelling percentage with the difference of mouse left and right sides auricular concha weight.
3.2 experimental result
Each dosage group of solamargine all can alleviate Oleum Tiglii and cause the mice ear degree, reduces the swelling rate, has compared significant difference (p＜0.05, p＜0.01) with the blank group.Show that solamargine has anti-inflammatory action, the results are shown in Table 3.
Table 3 solamargine to the influence of Oleum Tiglii induced mice ear swelling (
)
| Group | Dosage (mg/kg) | Number of animals (only) | Auricular concha swelling degree (mg) | The swelling rate |
| Blank group acetylsalicylic acid group solamargine low dose group | - 110 0.06 | 10 10 10 | 30.21±6.11 17.34±4.39 ** 27.47±6.59 | 197.5±56.3 113.4±40.2 ** 168.7±51.7 |
| Solamargine low dose group solamargine low dose group | 0.12 0.24 | 10 10 | 25.19±5.27 21.38±5.26 ** | 140.5±46.1 * 120.5±42.8 ** |
Annotate: compare with the blank group,
*P＜0.05,
*P＜0.01.
In sum, the injection solamargine is to S
180, Hep liver cancer transplanted tumor has stronger antitumor action, the injection solamargine also has certain anti-inflammatory action simultaneously.
(2) toxicological study acute toxicity test
Through acute toxicity test: intravenous administration of injection solamargine, LD
1Dosage is 20mg/kg, and being folded to the people is 2mg/kg.
Claims (10)
1. a plant alkaloid extract is characterized in that, it is from black nightshade herb and fruit, and perhaps Fruit of yellowfruit nightshade perhaps extracts in the Australian eggplant, and its extracting method step is as follows:
(1) gets black nightshade herb and fruit, perhaps Fruit of yellowfruit nightshade, perhaps Australian eggplant adds the 50-90% ethanolic soln and extracts 2-4 time, and each 4-8 doubly measures, reclaim ethanol, be concentrated into relative density 1.10～1.25, add 0.01～0.1mol/l acidic solution of 1～4 times of amount in the concentrated solution, fully stir, refrigeration was filtered more than 12 hours; Filter residue washs 1～3 time with 0.01～0.1mol/l acidic solution, and merging filtrate and washings are with low-pole or middle polarity resin absorption; Perhaps,
Get black nightshade herb and fruit, perhaps Fruit of yellowfruit nightshade, perhaps Australian eggplant, with 0.01～0.1mol/l acidic solution dipping 2～4 times, 4～8 times of each consumptions, merge steeping fluid, with low-pole or middle polarity resin absorption at each 6～24 hours time;
(2) first water washes resin column/bed to effluent liquid neutrality after the resin absorption, colourless with alkaline ethanol solution flushing resin column/bed again to effluent liquid, perhaps colourless with basic solution flushing resin column/bed earlier, colourless to contain alcohol amount 5%～40% ethanol liquid flushing resin column/bed again to effluent liquid to effluent liquid; Discard washing fluid, carry out wash-out with 50-80% ethanol again, show colourless, collect elutriant, reclaim ethanol, transfer pH value to 8-10, leave standstill refrigeration, collecting precipitation with basic solution to there not being the alcohol flavor to elutriant;
(3) get the precipitation drying after, with 20～25 times of amount organic solvent dissolutions, add the chromatographic silica gel of 0.5～4 times of amount of precipitation capacity, mix thoroughly, evaporate to dryness is got the silica gel of mixing behind the sample, column chromatography for separation, chromatography column is adorned post, organic solvent wash-out with the organic solvent wet method, elutriant is collected in gradation, thin-layer chromatography detects, and merges the elutriant of thin-layer chromatography detection and solamargine sample point position consistency, reclaims organic solvent, evaporate to dryness, promptly;
Perhaps,
(3) get the precipitation drying after, add the bigger organic solvent of polarity of 5～20 times of amounts, stirred 10～30 minutes, leave standstill, filter, filtrate adds the low polar organic solvent of 2～6 times of amounts, stir, refining, filter, reclaim organic solvent, add the recrystallization organic solvent, carry out recrystallization, collect crystallization, promptly.
2. a kind of plant alkaloid extract according to claim 1 is characterized in that the acidic solution described in the step (1) is selected from acetic acid, hydrochloric acid, sulfuric acid or citric acid solution; Basic solution described in the step (2) is selected from sodium hydroxide, calcium hydroxide, potassium hydroxide, magnesium hydroxide or the ammonia soln that concentration is 0.05-1.5mol/l, described alkaline ethanol solution is to contain the alkaline ethanol solution that is selected from sodium hydroxide, calcium hydroxide, potassium hydroxide, magnesium hydroxide or ammoniacal liquor that concentration is 0.05-0.15mol/l, and determining alcohol is 5%～40%.
3. a kind of plant alkaloid extract according to claim 1 is characterized in that the dissolving described in the step (3) is selected from methyl alcohol, dehydrated alcohol, acetone, Virahol or propylene glycol with organic solvent; Described column chromatography wash-out is selected from the mixed phase that in methyl alcohol, dehydrated alcohol, acetone, propyl carbinol, chloroform, the ethyl acetate or 2-3 item form with arbitrary proportion with organic solvent; The organic solvent that described polarity is bigger is selected from methyl alcohol, ethanol, propyl carbinol, acetone or two mixed phase wherein; Described low polar organic solvent is selected from chloroform, methylene dichloride, ethylene dichloride, ethyl acetate, toluene or sherwood oil; Described recrystallization is selected from 40%～80% methyl alcohol, 40%～70% ethanol, perhaps 20%～60% acetone with organic solvent.
4. one kind as any one described plant alkaloid extract formulation among the claim 1-3, it is characterized in that, it be the plant alkaloid extract and pharmaceutically the acceptable pharmaceutical carrier be mixed and made into the medicament of acceptable any formulation on the pharmaceutics.
5. a kind of plant alkaloid extract formulation according to claim 5 is characterized in that, in extract and pharmaceutical carrier mixing raw material, the weight percent content of extract is 0.1%-99.5%.
6. a kind of plant alkaloid extract formulation according to claim 5 is characterized in that, in extract and pharmaceutical carrier mixing raw material, the weight percent content of extract is 0.5%-95%.
7. a kind of plant alkaloid extract formulation according to claim 5 is characterized in that, described medicament is tablet or the capsule that extract adds weighting agent, disintegrating agent assembly; Or extract adds the slow releasing tablet or the capsule of weighting agent and hypromellose K4M assembly; Or extract is scattered in and obtains soft capsule in the oil phase; Or extract adds the injection of solubilizing agent or solubility promoter formation; Or extract is scattered in the injectable emulsion that obtains in the oil phase; Or extract adding pharmaceutical excipient is made suspension type injection liquid or freeze-drying injection powder pin.
8. a kind of plant alkaloid extract formulation according to claim 7 is characterized in that described weighting agent is selected from lactose, Microcrystalline Cellulose, dextrin, starch or calcium phosphate; Disintegrating agent is selected from hydroxypropylcellulose, sodium starch glycolate, polyvinylpolypyrrolidone or croscarmellose sodium; Also optional tackiness agent, lubricant or the wetting agent of adding when film-making agent and capsule; Described oil phase is selected from soybean oil, poly(oxyethylene glycol) 400, Oleum Gossypii semen, peanut oil, sesame oil, Semen Maydis oil or sweet oil; When the system soft capsule, also can add solubilizing agent or latent solvent and oxidation inhibitor etc.; The used solubilizing agent of injection is selected from Soxylat A 25-7 Viscotrol C, tween or pluronic F-68; Solubility promoter can be selected dilute hydrochloric acid, ornithine hydrochloride, arginine monohydrochloride, Citric Acid, acidic amino acid or other acid organism and bisalt thereof for use, and perhaps alcohols comprises ethanol, propylene glycol and N.F,USP MANNITOL.
9. a kind of plant alkaloid extract formulation according to claim 7, it is characterized in that, the preparation method of described suspension type injection liquid is, with extract and Polysorbate 80 mix grind after, be dissolved into the aqueous solution of phosphoric acid potassium dihydrogen, dipotassium hydrogen phosphate, nipagin esters and Xylo-Mucine, make through grinding.
10. the application of any one described a kind of plant alkaloid extract formulation in the medicine of preparation treatment liver cancer, lung cancer and other tumours among any one described a kind of plant alkaloid extract or the claim 4-9 among the claim 1-3, and the application in the preparation treatment inflammation medicine.
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| CN102000028A (en) * | 2010-11-03 | 2011-04-06 | 江苏康缘药业股份有限公司 | Solamargine liposome, preparation method and usage thereof |
| WO2011026637A3 (en) * | 2009-09-07 | 2011-04-28 | Thomas Kallimopoulos | Methods for isolating alkaloids from plants |
| CN103099891A (en) * | 2011-11-09 | 2013-05-15 | 安康市天源植物提取有限公司 | Preparation method of belladonna extract |
| CN104861024A (en) * | 2015-04-03 | 2015-08-26 | 上海交通大学 | Method for preparing beta-sitosterol linoleate from solanum nigrum plant |
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| CN100333753C (en) * | 2005-11-17 | 2007-08-29 | 刘阳 | Extraction technology of plant total alkaloid extract having anti tumour action and its preparation |
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| WO2011026637A3 (en) * | 2009-09-07 | 2011-04-28 | Thomas Kallimopoulos | Methods for isolating alkaloids from plants |
| CN102000028A (en) * | 2010-11-03 | 2011-04-06 | 江苏康缘药业股份有限公司 | Solamargine liposome, preparation method and usage thereof |
| CN102000028B (en) * | 2010-11-03 | 2012-09-05 | 江苏康缘药业股份有限公司 | Solamargine liposome, preparation method and usage thereof |
| CN103099891A (en) * | 2011-11-09 | 2013-05-15 | 安康市天源植物提取有限公司 | Preparation method of belladonna extract |
| CN104861024A (en) * | 2015-04-03 | 2015-08-26 | 上海交通大学 | Method for preparing beta-sitosterol linoleate from solanum nigrum plant |
| CN104861024B (en) * | 2015-04-03 | 2017-06-13 | 上海交通大学 | The method that Sitosterolum linoleate is prepared from black nightshade plant |
| CN106509579A (en) * | 2016-11-24 | 2017-03-22 | 张锦超 | Preparation method for nigrum fruit products |
| CN106509579B (en) * | 2016-11-24 | 2017-06-30 | 张锦超 | A kind of preparation method of black nightshade fruit product |
| CN108324798A (en) * | 2018-05-10 | 2018-07-27 | 李明慧 | A kind of converted products and purposes of S. photeinocarpum |
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