CN101450144A - Solanum dulcamara total-saponin extract and preparation method and use thereof - Google Patents
Solanum dulcamara total-saponin extract and preparation method and use thereof Download PDFInfo
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Abstract
The invention provides a solanum dulcamara total saponins extract containing 10-90% of solanum dulcamara total saponins by weight, which is a total steroidal saponins made from traditional Chinese medicine solanum dulcamara by process of extracting, separating and refining. Animal tests shows that: the exact has substantial effects of anti-inflammatory, anti-tumor, helicobacter pylori inhibition, gastric mucosa protection and keratinocyte proliferation inhibition, can be used in the preparation of medicaments for treating gastritis, stomach and duodenal ulcer stomach cancer and psoriasis.
Description
Technical field:
The present invention relates to Chinese medicine, be specifically related to a kind of extract and preparation method thereof of Chinese medicine and purposes in preparation treatment gastritis, gastric duodenal ulcer, psoriatic medicine.
Technical background:
The Chinese medicine Herba Solani Lyrati beginning is stated from Shennong's Herbal, classifies as top grade.Another name is also arranged: Herba Solani Lyrati, Herba Solani Lyrati, PAIFENGTENG etc.Herba Solani Lyrati has recorded in one one of Pharmacopoeia of the People's Republic of China version in 1977.Herba Solani Lyrati is a plant of Solanaceae, aboundresources, and its sweet in the mouth, hardship, cold in nature.Heat clearing away arranged, detoxify, dispel the wind, effect such as dampness removing, blood stasis dispelling.Be used for the treatment of jaundice due to damp-heat, rheumatic arthritis, leukorrhagia, edema, gonorrhea, erysipelas, furuncle, carcinoma.Modern study shows that the main chemical compositions in the Herba Solani Lyrati is steroidal saponin and Glycoalkaloids.Its water extract has antitumor action, and extracting the neutral steroidal saponin that obtains from its congener Fructus Solani dulcamarae then has the antifungal effect.CN1634295A has announced a kind of preparation method with Herba Solani Lyrati total alkaloids steroidal of antiinflammation, and CN1943676A has announced a kind of method for making of Lyrate nightshade gelatin and in the application aspect treatment eczema and the dermatomycosis.But,, there is no relevant record or report at present about Solanum dulcamara total-saponin and in the purposes for preparing treatment gastritis, gastric duodenal ulcer, gastric cancer, psoriatic medicine.
Gastritis comprises multiple stomach lining inflammation, can be divided into acute gastritis and chronic gastritis, and chronic gastritis can be divided into superficial gastritis, atrophic gastritis, intestinalization is living and typical hypertrophy.Gastritis may develop into gastric cancer, and especially atrophic gastritis, intestinalization are given birth to and typical proliferative phase.Gastritis mainly adopts the symptomatic treatment method, comprises eradicate helicobacter pylori, protection gastric mucosa etc.Peptic ulcer mainly refers to betide the duodenal chronic ulcer of harmonization of the stomach, and main the employing pressed down acid, protection gastric mucosa and anti-helicobacter pylori treatment.
Psoriasis be a kind of be the chronic dermatosis of feature with hyperproliferative epidermal, companion parakeratosis and corium lymphocytic infiltration, relate to the natural immunity, angiogenesis, t cell activation and excessive to a plurality of links such as skin, keratinocyte hypertrophy, main medicine has corticosteroids, retinoid, vitamin D-derivatives etc., though show certain short term efficacy, can not reach curative effect.
Summary of the invention
The technical problem to be solved in the present invention is to study a kind of Herba Solani Lyrati total glycosides extractive and preparation method thereof, and it is in the purposes of preparation treatment gastritis, gastric duodenal ulcer, gastric cancer, psoriatic medicine
The invention provides a kind of Herba Solani Lyrati total glycosides extractive, it contains percentage by weight is 10%～90% Solanum dulcamara total-saponin, more preferably its to contain percentage by weight be 35%～90% Solanum dulcamara total-saponin.
Solanum dulcamara total-saponin of the present invention, contain the steroidal saponin of one or more following structures:
Dulcain B
R＝3-O-α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→3)]-β-D-glucuronopyranoside
Dulcain C
R＝3-O-β-D-Glucopyranosyl-(1→3)-β-D-galactopyranoside
δ-solamarine
R＝3-O-α-L-Rhamnopyranosyl-(1→2)-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranoside
Dioscin.
Another object of the present invention has provided the preparation method of above-mentioned Solanum dulcamara total-saponin, and this method comprises the following steps:
1, extract:
After the pulverizing of crude drug process, water, methanol or ethanol, or aqueous alcohols, consumption is 4-10 a times of medical material weight, and room temperature or heating (comprising backflow) were extracted 1～3 hour, can repeat to extract 1～3 time, elimination medicinal residues then, filtrate is concentrated under normal pressure or decompression does not have the alcohol flavor, obtains concentrated solution, stand for standby use;
Wherein, extracting with solvent is good with 50%～95% ethanol, and the ethanol consumption is 4～10 times of medical material amount, and 6 times is good.
2, separate:
Said extracted liquid prepares Solanum dulcamara total-saponin by separation, and separation method can adopt one of following two kinds of methods:
(1) n-butanol extraction: its preparation process is that concentrated solution extracts 1-6 time (V/V) with 0.5 times of amount to 3 of n-butyl alcohol times amount, after the extraction, and evaporate to dryness, get extractum, gained extractum adds the petroleum ether that 1-6 doubly measures again, discards cleaning mixture, and solid drying gets Solanum dulcamara total-saponin extract;
(2) macroporous resin chromatography: its preparation process is that concentrated solution is with times 1-6 (V/V) defat of amount extraction of 0.5 times of amount to 3 of petroleum ether, resin column on the gained water layer, with water and rare alcohol (determining alcohol＜30%) washing, reuse 50-95% ethanol elution, collect eluent, concentrate, concentrated solution is drying to obtain Solanum dulcamara total-saponin;
The Solanum dulcamara total-saponin extract that said method makes can further be made with extra care purification to improve total saponin content.Concentrated solution is with 0.5 times of amount to 3 of n-butyl alcohol times 1-6 time (V/V) of amount extraction described in above-mentioned b, butanol extraction liquid is again with acid (hydrochloric acid solution of 0.1%-1%), alkali (sodium hydroxide of 0.1%-1% or sodium carbonate liquor) washing, and to be washed to neutrality, concentrate, drying can obtain the higher Solanum dulcamara total-saponin extract that gets of content.
By the Solanum dulcamara total-saponin of method for preparing, be black or yellow solid, can measure its total glycosides content with ultraviolet spectrophotometry is 10%～90%, preferably its total glycosides content is 35%～90%.
Another purpose of the present invention has provided the application of above-mentioned Herba Solani Lyrati total glycosides extractive in pharmacy.
The present invention will carry out a series of pharmacologically active screening and assessment with the Herba Solani Lyrati total glycosides extractive that said method makes, and the result shows that it has significant antiinflammatory, protection gastric mucosa, suppresses pharmacological actions such as helicobacter pylori, anti-stomach cancer cell propagation, anti-keratinocyte propagation.
1. the Herba Solani Lyrati total glycosides is to the protective effect of gastric mucosa of rat
Material: 50 of Wistar rats, body weight 160-180g, male, available from: experimental animal center, Chinese Academy of Sciences Shanghai, credit number: SCXK (Shanghai) 2003-0003.Given the test agent: press the preparation of embodiment method; Cimetidine (Cimetidine), lot number: 050801, available from: Heng Shan, Shanghai pharmaceutcal corporation, Ltd
Method: animal is according to the body weight random packet, and Herba Solani Lyrati total glycosides or positive control cimetidine are all used the 0.5%CMC suspendible, the oral administration gavage administration, and for three days on end, blank gives equal volume blank medium (0.5%CMC).Fasting is 24 hours before the last administration, gavages (p.o) dehydrated alcohol 1ml for every animal in 1 hour after the administration, puts to death animal after 1 hour, dissects and gets full stomach with the vernier caliper measurement ulcer index.
The result:
The Herba Solani Lyrati total glycosides causes the effect of rat acute gastric injury model to ethanol
*: compare P＜0.01 with the Vehicle group
The result shows: can cause acute erosive inflammation for rats gavaged ethanol, show as mucosal ulcer, hemorrhage equivalent damage symptom.The Herba Solani Lyrati total glycosides has obvious protective effect to rat pipe film injury due to the ethanol.
2. the Herba Solani Lyrati total glycosides is to the protective effect of mice gastric mucosa
Material: 32 of ICR male mices, 20-22g, experimental animal center, Chinese Academy of Sciences Shanghai provides; Herba Solani Lyrati total glycosides sample: press the preparation of embodiment method.
Method: animal adapts to 1-2 days.According to the body weight random packet, fasting is irritated stomach and is given Herba Solani Lyrati total glycosides or cimetidine after 48 hours, blank gives equal-volume solvent (0.5%CMC), gave every animal p.o dehydrated alcohol 0.1ml after the administration in 1 hour, put to death animal after 1 hour, dissection is got full stomach with formalin fixed, and the counting petechia is a ulcer index.
The result:
The Herba Solani Lyrati total glycosides causes the effect of chmice acute gastric injury model to ethanol
*: compare P＜0.01 with the Vehicle group;
The result shows that the Herba Solani Lyrati total glycosides has obvious inhibitory action to ethanol induced mice gastric mucosa injury (acute erosive inflammation).
3. the Herba Solani Lyrati total glycosides is to the inhibitory action of helicobacter pylori
Adopt flat band method, investigated the inhibitory action of Herba Solani Lyrati total glycosides to helicobacter pylori (H.Pylori), the result shows:
The result shows that the Herba Solani Lyrati total glycosides has remarkable inhibitory action to helicobacter pylori, minimal inhibitory concentration stomach 256 μ g/ml.
4. the inhibitory action of rat paw edema due to the Herba Solani Lyrati total glycosides on Carrageenan
Material: 40 of SD rats, male, body weight 130-150g, Shanghai Si Laike laboratory animal responsibility company limited provides the quality certification: SCXK (Shanghai) 2003-0003.Given the test agent: press the preparation of embodiment method.
Method: animal is divided into 4 groups at random, i.e. model contrast, positive control, given the test agent administration high dose group, low dose group, given the test agent 0.5%CMC suspendible, oral administration.Positive control-dexamethasone is a drug administration by injection.
Each treated animal is administered once equal every day, continuous 4 days, after the last administration 1 hour, inject 1% carrageenin and cause inflammation, cause scorching before earlier with drainage survey respectively organize the right back sufficient sole of the foot of rat volume once, cause inflammation at the right back sufficient sole of the foot middle part subcutaneous injection 1% carrageenin 0.1ml of rat then, measured sufficient sole of the foot volume once every one hour later on, rat cause scorching before and the difference that causes the right back sufficient sole of the foot volume of scorching back different time be the swelling value, calculate swelling rate and suppression ratio, compare with t method of inspection and model control group.
The result:
The given the test agent on Carrageenan causes the influence (X ± SD) of rat inflammation
*P＜0.05,
*Compare with model control group P＜0.01, and " 0 " expression causes scorching preceding measurement result, is swelling rate (%) in (), is suppression ratio in []..
The result shows that rat paw edema has remarkable inhibitory action due to the Herba Solani Lyrati total glycosides on Carrageenan, shows that the Herba Solani Lyrati total glycosides has antiinflammatory action.
5. medicine forms the influence of experiment to Mus tail epidermis granular cell
Material: the ICR mice, 26, male and female half and half are provided by Shanghai Slac Experimental Animal Co., Ltd., credit number: SCXK (Shanghai) 2003-0003; All-trans-retinoic acid (ATRA): auspicious chemical industry company limited product, the lot number: 070429 of reaching in Zhengjiang City; Herba Solani Lyrati total glycosides: press the preparation of embodiment method; Preparation: be subjected to reagent thing and positive control ATRA all with after a small amount of DMS0 dissolving, make liniment with ethanol dilution, concentration is 0.5%; Be coated with Mus tail administering mode outside adopting, once a day, be 14 days the course of treatment.
Method: with the mice random packet, male and female half and half.Each treated animal is smeared afterbody with relative medicine respectively, and blank group (Vehicle) gives the blank solvent of equal capacity, and every day 1 time, coating was put to death mice next day after 14 days continuously.Get the 1.8-4cm place epidermis apart from root of the tail portion, after 10% formaldehyde fixed, carry out tissue slice and HE dyeing, optical microscope is observed down.Observation index: the scale epidermis between 1. all two follicular orifices has the granular layer person who is arranged in rows to count the scale that granular layer forms, and counts the scale number that granular layer is arranged in per 100 scales; 2. count granular cell number in 100 scales of Mus tail epidermis, granular cell is formed by flat in the epidermis or rhombus cell, is full of dark basophilia in the endochylema, and is thick, and HE dyeing is the transparent cutin granule of navy blue.Each group data is compared, and take statistics is handled.
The result:
Mouse tail scale granular layer of epidermis is formed by the reagent thing and the influence of granular cell number (x ± s)
Annotate: compare with the Vehicle group,
*Be p＜0.05,
*Be p＜0.01
6, Solanum dulcamara total-saponin is to the antiproliferative effect of people's immortalization epidermis cell strain HaCaT cell
Material and method: HACAT cell: available from Chinese Academy of Sciences's cellular biochemical institute.All-trans-retinoic acid (ATRA): auspicious chemical industry company limited product, the lot number: 070429 of reaching in Zhengjiang City; Herba Solani Lyrati total glycosides: press the preparation of embodiment method; The HACAT cell that will be in exponential phase is with 3 * 10
4/ ml is inoculated in and contains 10% hyclone (GIBCO); 1 * 10
5The U.L-1 penicillin, 1 * 10
5The RPMI1640 culture medium of ug.L-1 streptomycin is at 37 ℃, 5%CO
2Cultivated 72 hours under the condition, srb assay detects the inhibition activity that is subjected to reagent thing and control drug on cell proliferation.
The result:
Solanum dulcamara total-saponin is to the inhibitory action of HACAT cell proliferation
The keratinocyte paraplasm is one of psoriatic main diseases variation of science, and result of study shows that the Herba Solani Lyrati saponin has remarkable inhibiting activity to keratinocyte (HACAT), and prompting can be used for psoriatic treatment.
7. Solanum dulcamara total-saponin is to the antiproliferative effect of stomach cancer cell
Material and method: BGC-823 cell: available from Chinese Academy of Sciences's cellular biochemical institute.Herba Solani Lyrati total glycosides: press the preparation of embodiment method; The BGC-823 cell that will be in exponential phase is with 3 * 10
4/ ml is inoculated in and contains 10% hyclone (GIBCO); 1 * 10
5The U.L-1 penicillin,
The RPMI1640 culture medium of ug.L-1 streptomycin is at 37 ℃, 5%CO
2Cultivated 72 hours under the condition, srb assay detects the inhibition activity that is subjected to reagent thing on cell proliferation.
The result:
Solanum dulcamara total-saponin has obvious inhibitory action, IC50:18.84 μ g/ml to BGC-823 cells in vitro propagation.
Above-mentioned studies show that: the Herba Solani Lyrati total glycosides has significant antiinflammatory, protection gastric mucosa, suppresses pharmacologically actives such as helicobacter pylori, anti-stomach cancer cell propagation, anti-keratinocyte propagation, can be used for preparation treatment gastritis, gastric duodenal ulcer, gastric cancer, psoriatic medicine.
This Herba Solani Lyrati total glycosides by making oral or non-oral formulation according to a conventional method with pharmaceutic adjuvant, is used for gastritis, gastric duodenal ulcer, gastric cancer, psoriatic treatment as active component.Be preferably oral solid formulation especially, can make various dosage forms such as tablet, capsule, granule, pill according to a conventional method with adjuvant.
The specific embodiment of form is described in further detail foregoing of the present invention again by the following examples.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following example.All technology that realizes based on foregoing of the present invention all belong to scope of the present invention.
The specific embodiment
Embodiment 1:
Herba Solani Lyrati 1000g adds 75% ethanol 6L, heating and refluxing extraction 1 hour, and filtering residue is again with 75% ethanol extraction twice, each 6L, reflux 1 hour is filtered merging filtrate.Filtrate is concentrated to a certain degree, and with n-butanol extraction (500ml * 4), n-butyl alcohol is to wash the back concentrate drying, and the gained solid filters with petroleum ether, reclaims solid, gets the Herba Solani Lyrati total glycosides, and yield is 3.6%, and recording its total glycosides content is 49.4%
Embodiment 2:
Herba Solani Lyrati 1500g adds 50% ethanol 7L, heating and refluxing extraction 1 hour, and filtering residue is again with 50% ethanol extraction twice, each 5L, reflux 1 hour is filtered merging filtrate.Filtrate is concentrated to a certain degree, with petroleum ether extraction, and after the extraction, D101 macroporous resin on the water, water, 20% washing with alcohol are then used 50% ethanol elution successively, and the eluent concentrate drying gets the Herba Solani Lyrati total glycosides, and recording its total glycosides content is 65.3%
Embodiment 3:
Herba Solani Lyrati 1500g adds pure water 7L, heating and refluxing extraction 1 hour, and filtering residue extracts twice with pure water again, each 5L, reflux 1 hour is filtered merging filtrate.Filtrate is concentrated to a certain degree, with petroleum ether extraction, after the extraction, D101 macroporous resin on the water with water-20% ethanol-50% ethanol-95% ethanol elution, is collected 50% ethanol elution position successively, be concentrated into and do not have the alcohol flavor, concentrated, dry, promptly get Solanum dulcamara total-saponin extract, recording its total glycosides content is 59%.
Claims (7)
1, a kind of Solanum dulcamara total-saponin extract is characterized in that it is 10%～90% Solanum dulcamara total-saponin that this extract contains percentage by weight, is preferably the total saponins of 35%-90%.
2, Solanum dulcamara total-saponin extract according to claim 1 is characterized in that this extract has one or more in the steroidal saponin of following structure:
Dulcain B
Dulcain C
δ-solamarine
Dioscin.
3, a kind of preparation method of Solanum dulcamara total-saponin extract as claimed in claim 1 is characterized in that this method comprises the following steps:
(1) extract:
With crude drug through after pulverizing, water, methanol or ethanol, or aqueous alcohol, room temperature or heating or reflux, extract, 1～3 hour can repeat to extract 1～3 time, elimination medicinal residues then, filtrate is concentrated under normal pressure or decompression does not have the alcohol flavor, obtains concentrated solution, stand for standby use.
(2) separate:
A, n-butanol extraction: its preparation process is that concentrated solution is with 0.5 times of amount to 3 of n-butyl alcohol times amount V/V extraction 1-6 time, after the extraction, evaporate to dryness gets extractum, and gained extractum adds the petroleum ether V/V washing that 1-6 doubly measures again, discard cleaning mixture, solid drying gets Solanum dulcamara total-saponin extract; Or
B, the macroporous resin chromatography: its preparation process is that concentrated solution is with times 1-6 the defat of amount V/V extraction of 0.5 times of amount to 3 of petroleum ether, resin column on the gained water layer, with water and rare pure determining alcohol＜30% washing, reuse 50-95% ethanol elution, collect eluent, concentrate, concentrated solution is drying to obtain Solanum dulcamara total-saponin.
4, the preparation method of Solanum dulcamara total-saponin extract according to claim 3 is characterized in that solvent is water, methanol or ethanol or aqueous alcohol described in this method step (1) extraction; Preferred 50%～95% ethanol; Solvent load is 4～10 times of medical material weight, preferred 6 times.
5, the preparation method of Solanum dulcamara total-saponin extract according to claim 3 is characterized in that the described resin column of this method step (2) is D101 macroporous resin column, AB-8 or HPD100 macroporous adsorbent resin.
6, the purposes of a kind of Solanum dulcamara total-saponin extract according to claim 1 in preparation treatment gastritis, gastric duodenal ulcer, gastric cancer or psoriasis medicine.
7,, it is characterized in that described medicine is oral, injection or the percutaneous drug administration preparation that Solanum dulcamara total-saponin extract and medical dressing are made according to the purposes of the described Solanum dulcamara total-saponin extract of claim 6 in pharmacy.
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| CN102133298A (en) * | 2010-12-03 | 2011-07-27 | 沈阳药科大学 | Two-aqueous-phase method for extracting total saponin from bittersweet herb |
| CN102319319A (en) * | 2011-09-15 | 2012-01-18 | 中国中医科学院西苑医院 | Solanum lyratum extract, and anticancer medicine comprising solanum lyratum extract |
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| CN110327354A (en) * | 2019-08-12 | 2019-10-15 | 北京市中医研究所 | Application of the diosgenin in curing psoriasis and prevention |
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| CN102133298A (en) * | 2010-12-03 | 2011-07-27 | 沈阳药科大学 | Two-aqueous-phase method for extracting total saponin from bittersweet herb |
| CN102319319A (en) * | 2011-09-15 | 2012-01-18 | 中国中医科学院西苑医院 | Solanum lyratum extract, and anticancer medicine comprising solanum lyratum extract |
| CN102319319B (en) * | 2011-09-15 | 2013-05-08 | 中国中医科学院西苑医院 | Solanum lyratum extract, and anticancer medicine comprising solanum lyratum extract |
| CN105726554B (en) * | 2016-02-02 | 2019-05-10 | 大连医科大学 | Dioscin is preparing the application in acute gastrointestinal damage protection drug |
| CN105726554A (en) * | 2016-02-02 | 2016-07-06 | 大连医科大学 | Application of dioscin in preparation of acute gastrointestinal injury protecting medicine |
| CN110327354A (en) * | 2019-08-12 | 2019-10-15 | 北京市中医研究所 | Application of the diosgenin in curing psoriasis and prevention |
| CN110327354B (en) * | 2019-08-12 | 2023-08-22 | 北京市中医研究所 | Application of diosgenin in treatment and prevention of psoriasis |
| CN113521262A (en) * | 2021-07-30 | 2021-10-22 | 浙江莱康生物工程有限公司 | Lysozyme preparation with anti-inflammatory effect |
| CN113521262B (en) * | 2021-07-30 | 2023-10-13 | 浙江莱康生物工程有限公司 | Lysozyme preparation with anti-inflammatory effect |
| CN115505022A (en) * | 2022-07-22 | 2022-12-23 | 上海市中医医院 | Alkaloid glycoside and application thereof |
| CN115505022B (en) * | 2022-07-22 | 2023-11-17 | 上海市中医医院 | Alkaloid glycoside and application thereof |
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