CN1158088C - Water extracted and alcohol precipitated matter of subprostrate sophora and its application in medicine - Google Patents
Water extracted and alcohol precipitated matter of subprostrate sophora and its application in medicine Download PDFInfo
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- CN1158088C CN1158088C CNB001140566A CN00114056A CN1158088C CN 1158088 C CN1158088 C CN 1158088C CN B001140566 A CNB001140566 A CN B001140566A CN 00114056 A CN00114056 A CN 00114056A CN 1158088 C CN1158088 C CN 1158088C
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- radix sophorae
- sophorae tonkinensis
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Abstract
The present invention relates to a water extracted and alcohol precipitated substance of vietnamese sophora root, which is prepared by the following steps: soaking vietnamese sophora root by water, extracting later through heating, and then extracting by hot water for 0 to 3 times; filtering, merging the water extracting solution, concentrating, adding alcohol, and then taking the precipitated part in the alcohol so as to obtain the substance. The substance has the effects of protecting the liver, regulating immunity and resisting tumors and can be used for preparing medicines for the purposes, and has low toxicity.
Description
The present invention relates to a kind of extraction of effective ingredient of Chinese medicine Radix Sophorae Tonkinensis and the application in pharmacy thereof.
Radix Sophorae Tonkinensis is the root of leguminous plant sophora subprostrata (root of subprostrate sophora) sophrora subprostrataChun et T.Chen.Mainly originate in province's nature and flavor bitter colds such as Guangxi, Guangdong, Guizhou and return lung meridian.Pharmacopeia record Radix Sophorae Tonkinensis has heat-clearing and toxic substances removing, throat, and the effect of loosing and swelling and ache, the main component of Radix Sophorae Tonkinensis is that alkaloids comprises matrine, oxymatrine, anagyrines etc., flavonoid comprise that Chinese scholartree is fixed, Chinese scholartree ketone, sophoradochromene, Chinese scholartree promise chromene.The utilization of modern study Radix Sophorae Tonkinensis has the report of treatment hepatitis, and the alkaloid of useful Radix Sophorae Tonkinensis partly is used for the treatment of the report of hepatitis.The composition of Radix Sophorae Tonkinensis water extract-alcohol precipitation part is mainly polysaccharide, and the extracting method of the xyloglucan in the Radix Sophorae Tonkinensis has report, but unmanned up to now with this polysaccharide component as medicinal part, less than report about its medical value.Zoopery proof Radix Sophorae Tonkinensis is not only water-soluble but also be dissolved in alcoholic acid total extract (alkaloid and flavonoid are main) to mouse stomach (25g/kg), symptom such as respiration inhibition, ultra-large type are trembled, tic occurs even cause animal dead, thereby Radix Sophorae Tonkinensis was not only water-soluble but also be dissolved in alcoholic acid total extract and have certain toxicity.
Purpose of the present invention is exactly for the report at the medical value of unmatchful its water extract-alcohol precipitation part in present Radix Sophorae Tonkinensis effective ingredient medicinal, provide have low toxicity, Radix Sophorae Tonkinensis extract (water extracting alcohol hypostasis), its extracting method and the application in pharmacy thereof that therapeutic effect is good.
The present invention realizes like this.
Get Radix Sophorae Tonkinensis, the post-heating that is soaked in water extracts, and behind 0～3 hot water extraction, filters again, merges the water extract, concentrates, adds ethanol, gets the precipitate with ethanol part promptly.
Radix Sophorae Tonkinensis water extracting alcohol hypostasis of the present invention is meant can be water-soluble, especially hot water and in ethanol insoluble sedimentary part.Never this part is used as medicinal part in the report in the past.
The extracting mode of Radix Sophorae Tonkinensis water extracting alcohol hypostasis of the present invention also can adopt earlier and discard with ethanol extraction Radix Sophorae Tonkinensis alcohol soluble components, gets shallow slag, and water is carried promptly again.
Behind the Radix Sophorae Tonkinensis hot water extraction of the present invention, filtration is filtered into good with filtered while hot or insulation.
After Radix Sophorae Tonkinensis water extract of the present invention concentrated, adding ethanol, to make content be 80%.
Above-mentioned Radix Sophorae Tonkinensis water extracting alcohol hypostasis through anthrone method polysaccharide qualitative experiment, is positive, and through phenol-sulfuric acid process assay, shows higher polyoses content, shows that Radix Sophorae Tonkinensis water extracting alcohol hypostasis is based on polysaccharide.
The acute toxicity test of Radix Sophorae Tonkinensis water extracting alcohol hypostasis is: to the chmice acute toxicity test, measure mouse death rate with karber's method with the Radix Sophorae Tonkinensis polysaccharide, change because its toxicity is very little and do the maximum tolerated dose experiment.The result: Radix Sophorae Tonkinensis water extracting alcohol hypostasis acute toxicity is very low, with being equivalent to crude drug 166.8g/kg (for irritating the Cmax of stomach), observes seven days, and none example is dead, and this dosage is equivalent to 500 times of clinical ordinary person's consumption.
Conclusion: the safety of Radix Sophorae Tonkinensis water extracting alcohol hypostasis is good, is the low Chinese medicine safe in utilization of a toxicity.
Radix Sophorae Tonkinensis water extracting alcohol hypostasis shows the experiment of the protective effect of acute liver damage: Radix Sophorae Tonkinensis water extracting alcohol hypostasis is with resisting CCl
4Due to the rat acute liver injury model, detect the changes of contents of AST, ALT, TBlL in rats death rate and the serum.The result shows that Radix Sophorae Tonkinensis water extracting alcohol hypostasis significantly reduces Rats with Acute Liver Injury mortality rate and serum AST, ALT, TBlL content, shows that Radix Sophorae Tonkinensis water extracting alcohol hypostasis has significant liver protection effect.
Material
1. animal SD rat is 50, male and female half and half, body weight 200g～250g.
2. main medicine and reagent: Radix Sophorae Tonkinensis water extracting alcohol hypostasis, hepatitis spirit, diammonium glycyrrhizinate, CCl
4(analytical pure), edible peanut oil.
3. each method for preparation of drug:
The hepatitis spirit is used after being diluted to 10 times:
After being diluted to 5ml solution, uses diammonium glycyrrhizinate every (50mg);
Radix Sophorae Tonkinensis water extracting alcohol hypostasis 3g/kg
4. main equipment Hitachi7170A fully-automatic analyzer (Hitachi)
Method
Animal experiment method: rat is divided into 5 groups at random by table 1.Press table 2 successive administration 9 times (blank group and model group are given the normal saline of equivalent), each 24h at interval.Get the preceding 18h of blood, to subcutaneous injection 50%CCl behind the rat neck
4Oleum Arachidis hypogaeae semen mixed solution 0.072ml/100g modeling (normal control group injection equivalent normal saline) is appended in modeling and is administered once, and the beginning fasting.Modeling 17h, each group is given corresponding medicine once more, and the 1h eye socket is got blood 2ml after the administration, the separation of serum censorship.
Observation item:
1. mortality analysis (seeing Table 1)
Table 1, rat grouping and dead result
Number of animals (only) death toll (only)
Group
Mortality rate
18h after male and female half and half modeling
Blank group 10 00
Model group 10 4 40%
Hepatitis spirit group 10 3 30%
Diammonium glycyrrhizinate group 10 3 30%
Radix Sophorae Tonkinensis water extracting alcohol hypostasis 10 1 10%
By experiment as seen, in experimental group positive controls and experimental group mortality rate all less than model control group.
Serum glutamic pyruvic transminase (ALT) enzyme process (reagent source: company is given birth to by Taiwan unit)
Serum glutamic oxalacetic transaminase (ALT) is with biuret method (reagent source: German Human company)
The result:
The variation that each organizes ALT in the acute liver damage, AST content the results are shown in Table 2.
AST, ALT in table 2 acute liver damage, TBlL changes of contents (x ± SD)
The group n Dose AST (ALT of μ/the L) (TBlL of μ/L)
Blank group 10 NS (ip/ irritates stomach) 226 ± 61
*64 ± 26*, 6.03 ± 4.42*
Model group 6 NS (ip/ irritates stomach) 1823 ± 467 1854 ± 941 11.31 ± 4.63
Hepatitis spirit group 7 0.028ml/100g (ip) 1668 ± 1406*** 3016 ± 1,792 11.11 ± 1.38
Diammonium glycyrrhizinate group 7 3.6ml/100g (filling stomach) 1072 ± 709**, 1872 ± 2377*** 8.56 ± 3.59
Radix Sophorae Tonkinensis water
Carry pure hypostasis 8 0.38ml/100g (filling stomach) 741 ± 548* △ ▲ 346 ± 328**, 6.49 ± 2.23* ▲
Annotate: relatively organize relatively △ P＞0.05 and diammonium glycyrrhizinate group comparison ▲ P＞0.05 with the hepatitis spirit in * P＜0.01 * * P＜0.05 * * * P＞0.05 with model
From then on showing visible Radix Sophorae Tonkinensis water extracting alcohol hypostasis can obviously reduce the AST in the blood, the content of ALT than model group, and significantly better than positive controls, so Radix Sophorae Tonkinensis water extracting alcohol hypostasis is to CCl
4The acute liver damage that is caused has significant protective effect.
Discuss
Model evaluation CCl
4Due to hepatocyte injury mechanism be, influence the metabolism of cytochrome oxidase in the hepatocyte microsome (P450), produce the active free radical CCl of Cl
3, the time and produce chlorine alkanisation and lipid peroxidation, destroy the membranous structure of cell, blocking protein is synthetic, secretion and other liver function, and directly toxic action is by CCl
3The lipid peroxidation that produces causes the active rising of liver plasma membrane, and hepatocyte injury also makes blood mesobilirubin content raise.Therefore, the mensuration of AST, ALT is to judge the part important indicator of hepatocyte injury, and general content is high more, and liver cell lesion is obvious more.Simultaneously, AST, ALT all belong in the non-specific cell functional enzyme just often the serum intensive amount are very low, and when the green cell membrane permeability increased, its activity increased sharply.ALT mainly is present in the liver pulp, because of enzymatic activity in the liver is high approximately 100 times than serum, as long as therefore 1% stem cell necrosis is arranged, the ALT in the serum is doubled, and it is that the most responsive liver function detects one of index.AST is the highest in levels and myocardial contents, and liver is second, and when hepatic injury, it secretes output is low than ALT, and therefore, the sensitivity of AST assaying reaction hepatocyte injury is low than ALT.
From the pathogen and pathology of tcm angle, generally believe, Fibrotic etiology and pathogenesis is " damp and hot residual poison is not to the greatest extent; stagnation of blood stasis and heat yin collaterals; stagnation of QI due to depression of the liver; spleen kidney yin yang-energy blood deficiency " after the hepatitis, the simulataneous insufficiency and excessive situation, wherein damp and hot poison of the stasis of blood and spleen deficiency of the kidney are the pathogenesis keys, and its pattern of syndrome is clinical, and in the majority (repeatedly clinical research finds that traditional Chinese medical science what is called " stagnant heat, stagnation of liver-QI, spleen kidney two lose " is whole with immune disorder, and degree of hepatic fibrosis is relevant, still adopt benefiting QI for activating blood circulation, 'Shugan Lidan ', removing liver heat and toxic substances, the prescription of dampness dampness removing is treated hepatic fibrosis.
Each group is at injection CCl in this experiment of drug evaluation
4After, wherein model group rats death rate is the highest, and serum AST, ALT raise the most obvious.Matched group and experimental group all can reduce above-mentioned various index (hepatitis spirit group ALT exception), but the experimental group drug effect is good than matched group, and the ALT effect is the most obvious.And the heavy dose of group effect of Radix Sophorae Tonkinensis water extracting alcohol hypostasis is better than small dose group, and visible Radix Sophorae Tonkinensis water extracting alcohol hypostasis has liver protection effect preferably.
Pharmacodynamic experiment shows that Radix Sophorae Tonkinensis water extracting alcohol hypostasis has immunomodulating and antineoplastic obviously acts on.
In a word, Radix Sophorae Tonkinensis water extracting alcohol hypostasis have significantly protect the liver, immunomodulating and antitumor action, can be used to prepare the medicine of such use, obviously be better than the alkaloidal effect of present Radix Sophorae Tonkinensis.Toxicity test shows, Radix Sophorae Tonkinensis water extracting alcohol hypostasis low toxicity was not only water-soluble but also be dissolved in alcoholic acid total extract toxicity and greatly reduce than Radix Sophorae Tonkinensis alkaloid and Radix Sophorae Tonkinensis.
Adult's oral dose is every day three times, to take the Radix Sophorae Tonkinensis water extracting alcohol hypostasis that is equivalent to 15 gram crude drugs at every turn.
The invention will be further described below in conjunction with embodiment.
Embodiment 1
Take by weighing the Radix Sophorae Tonkinensis 50g that shreds, spend the night with 12 multiple dose water loggings bubble, heating extraction 2h, water outlet liquid inclines, centrifugal while hot (3000r/min, 10min) medicinal residues continue to add 10 times, 8 multiple dose water are heating extraction 1.5h respectively, 1h, the water outlet liquid that inclines separately is centrifugal while hot, water liquid antigradient is concentrated into 1: 1 (the 1g crude drug is equivalent to 1ml), add 95% ethanol and make and contain alcohol amount and reach 80%, the stand at low temperature back of spending the night is centrifugal, and precipitation adds behind 1: 50 hot water dissolving centrifugal again, water intaking liquid adds 95% ethanol makes the alcohol amount reach 80%, centrifugal after stand at low temperature is spent the night, it is centrifugal again that precipitation adds the stirring of 95% ethanol, precipitation vacuum drying (40 ℃), get crude product Radix Sophorae Tonkinensis water extracting alcohol hypostasis, and weigh.
After the extract drying, granulate with food starch, make electuary.
Embodiment 2
Radix Sophorae Tonkinensis was used 10 times of water loggings bubble instead 12 hours, and medicinal residues continue with 8 times of water, 6 times of water heating extraction, extraction conditions all the other with embodiment 1.
The said extracted thing is made the preparation of targeting preparation organ liposome: phospholipid is dissolved in an amount of ether, adds water extracting alcohol hypostasis aqueous solution, make its ratio greatly about (aqueous solution: organic solvent=1: 3～1: 6).Carry out ultrasonic in short-term, up to forming stable W/O Emulsion.Remove organic solvent at reduction vaporization then, reach colloidal state after, other adds in a certain amount of phosphate buffer (PBS), rotation helps the gel on the wall to come off, and under reduced pressure continues evaporation, the centrifugal medicine that does not wrap into of removing then, after promptly getting liposome, embedding.
Embodiment 3
Radix Sophorae Tonkinensis spends the night with 8 times of water logging bubbles, heating extraction 3 hours, and filtered while hot discards medicinal residues.Filtrate is concentrated into 1: 1, adds ethanol, makes to contain the alcohol amount and reach 60%, and stand at low temperature is centrifugal, taking precipitate, vacuum drying.
Liposome-antibody formation immunoliposome of deriving
Get the Palmic acid N-Hydroxysuccinimide fat (NHSP) that is dissolved in dioxane, add an anti--HBs monoclonal antibody, 2% NaTDC, evenly, room temperature is placed 2h, and the SephadexG75 gel chromatography is removed surplus NaTDC and PBS, will add above-mentioned finger plastid in the Palmic acid antibody covalent cross-linked thing of gel chromatography purification, after the mixing, prepare immunoliposome with the Liposomate instrument, embedding.
Various embodiments of the present invention all have hepatoprotective, immunomodulating, antineoplastic action, and toxicity is low, Radix Sophorae Tonkinensis water extracting alcohol hypostasis of the present invention can be processed into electuary, solvent, capsule, oral liquid, durative action preparation, targeting preparation etc., the invention is not restricted to the foregoing description.
Claims (5)
1. Radix Sophorae Tonkinensis extract is characterized in that: it is the precipitate with ethanol part that adds the ethanol gained in Radix Sophorae Tonkinensis water extract; Or the water of the residue reuse water extraction gained after the Radix Sophorae Tonkinensis alcohol extraction put forward part.
2. the extracting method of Radix Sophorae Tonkinensis extract as claimed in claim 1 is characterized in that having the following steps successively: get Radix Sophorae Tonkinensis, the post-heating that is soaked in water extraction, behind 0～3 hot water extraction, filter again, merge the water extract, concentrate, add ethanol, get the precipitate with ethanol part promptly.
3. the extracting method of Radix Sophorae Tonkinensis extract according to claim 2 is characterized in that Radix Sophorae Tonkinensis hot water extraction after-filtration is that filtered while hot or insulation are filtered.
4. the extracting method of Radix Sophorae Tonkinensis extract according to claim 2, it is characterized in that Radix Sophorae Tonkinensis hot water extraction liquid concentrates after, add ethanol and make content reach 80%.
5. Radix Sophorae Tonkinensis extract as claimed in claim 1 is used to prepare the purposes of hepatinica.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB001140566A CN1158088C (en) | 2000-01-31 | 2000-01-31 | Water extracted and alcohol precipitated matter of subprostrate sophora and its application in medicine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB001140566A CN1158088C (en) | 2000-01-31 | 2000-01-31 | Water extracted and alcohol precipitated matter of subprostrate sophora and its application in medicine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1306854A CN1306854A (en) | 2001-08-08 |
| CN1158088C true CN1158088C (en) | 2004-07-21 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB001140566A Expired - Fee Related CN1158088C (en) | 2000-01-31 | 2000-01-31 | Water extracted and alcohol precipitated matter of subprostrate sophora and its application in medicine |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101040899B (en) * | 2006-03-20 | 2013-06-19 | 周亚伟 | Antineoplastic Chinese traditional compound and the preparation and the method for producing the same |
| CN1994364B (en) * | 2006-12-30 | 2010-08-25 | 湖南康普制药有限公司 | Subprostrate soplhor root extract refining process |
| CN103040919B (en) * | 2012-12-29 | 2015-04-15 | 黄家芬 | Method for extracting general flavone from red bean |
| CN106822228B (en) * | 2015-12-03 | 2021-05-04 | 上海中医药大学 | Subprostrate sophora polysaccharide effective part and preparation method thereof |
| CN106822229B (en) * | 2015-12-03 | 2020-06-26 | 上海中医药大学 | Application of subprostrate sophora polysaccharide effective part |
| CN107126457B (en) * | 2016-02-29 | 2020-10-30 | 上海中医药大学 | Application of subprostrate sophora polysaccharide effective part in preparing antitumor drug |
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2000
- 2000-01-31 CN CNB001140566A patent/CN1158088C/en not_active Expired - Fee Related
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