CN108976264A - A kind of kidney tonifying, benefiting essence-blood, radiation protection saussurea involucrata Herba Epimedii root of kirilow rhodiola Chinese medicine composition - Google Patents

A kind of kidney tonifying, benefiting essence-blood, radiation protection saussurea involucrata Herba Epimedii root of kirilow rhodiola Chinese medicine composition Download PDF

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CN108976264A
CN108976264A CN201810856507.5A CN201810856507A CN108976264A CN 108976264 A CN108976264 A CN 108976264A CN 201810856507 A CN201810856507 A CN 201810856507A CN 108976264 A CN108976264 A CN 108976264A
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extract
herba epimedii
blood
root
chinese medicine
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杨桂芹
王志标
刘霞
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Anhui Jin Yuan Pharmaceutical Co Ltd
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Anhui Jin Yuan Pharmaceutical Co Ltd
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Abstract

Disclose a kind of kidney tonifying, benefiting essence-blood, radiation protection saussurea involucrata Herba Epimedii root of kirilow rhodiola Chinese medicine composition, and extract glycoside substance method, the composition is the Chinese medicine composition comprising Shorthorned Epimedium P.E and gadol extract, the extract has particularly good raising testosterone secretion effect compared with general extraction mixture.The composition, which can be used for preparing, can enhance immune function of human body, antifatigue, radiation protection and anti-inflammatory and kidney tonifying, the drug of benefiting essence-blood, health care product or drink.

Description

A kind of kidney tonifying, benefiting essence-blood, radiation protection saussurea involucrata Herba Epimedii root of kirilow rhodiola Chinese medicine composition
Technical field
The present invention relates to extract and preparation technique field, in particular to a kind of kidney tonifying, benefiting essence-blood, radiation protection saussurea involucrata Herba Epimedii Root of kirilow rhodiola Chinese medicine composition, and glycoside substance method is extracted from Herba Epimedii using multistage column chromatography for separation, and be related to wrapping Composition containing Shorthorned Epimedium P.E and root of kirilow rhodiola derivative, in particular to a kind of raising immunity, antifatigue, radiation protection and Kidney tonifying, benefiting essence-blood composition, the composition especially includes the Chinese medicine composition of Shorthorned Epimedium P.E and gadol extract, The either Chinese medicine composition comprising Saussurea involucrata extract, Shorthorned Epimedium P.E and gadol extract.
Background technique
Herba Epimedii is traditional traditional tonic medicine in China, is initially recorded on Shennong's Herbal, with tonifying kidney and strengthening yang, by force Muscle reinforcing and bone strengthening, the curative effect of remove rheumatism.Epimedium herb is the dried leaf of Berberidaceae plant Herba Epimedii, and existing pharmacopeia is included shared The kinds such as Epimedium sagittatum, E. Pubescens and korean epimedium herb are more.Herba Epimedii has the function of kidney-replenishing, strengthening the bones and muscles, wind-damp dispelling Effect.Icariin is one of monomer ingredient, and having improves cardiovascular system, adjusts endocrine, strengthen immunity, sex hormone sample The effects of, while also having positive therapeutic effect in terms of antitumor, anti-liver.
Modern pharmacological studies have shown that Herba Epimedii can increase cardiovascular and cerebrovascular blood flow, promote hematopoiesis function, immune function and bone Metabolism, enhancing testosterone secretion are " kidney tonifying, benefiting essence-blood " effect, also have anti-aging, antitumor and other effects.
The kidney tonifying of Herba Epimedii and benefiting essence-blood effect and mechanism have also obtained extensive approval and research.In the whole nation that Li Mei writes In higher medical institution pharmacy class fourth round programming textbook " traditional Chinese medicine and pharmacy basis " (the 3rd edition [M] .2016), excessive sheep is explicitly pointed out The leaves of pulse plants has efficacies of tonifying kidney and invigorating yang, and pharmacology mainly enhances immune function, improves gonad function and anti-aging, is anti-inflammatory, anti-mistake It is quick etc..In " the TCM specialty knowledge " that state food pharmaceuticals administration general bureau licensed pharmacist qualification authentication central tissue writes (2015) the warming kidney and enhancing yang effect of Herba Epimedii and its extract (mainly flavonoids and glycosides compound) is also explicitly pointed out.It is opening " conventional Chinese medicine effect compares " (2013) that frightened lake is write have explicitly pointed out the kidney tonifying of Herba Epimedii and its extract, benefiting essence-blood is made With.
Recent researches discovery, the monomeric substances such as glycoside substance, Flavonoid substances and polysaccharose substance in Herba Epimedii, Improve blood vessel inner skin cell function and be better than tradition processing Herba Epimedii Chinese medicine especially, however their separation and Extraction has very big difficulty.
Saussurea involucrata is the rare rare Chinese medicine in China, have it is remote with history and civil medication basis abundant, The flavones and its glycosides that contain in saussurea involucrata, Phenylpropanoid Glycosides class, sesquiterpene lactone, cumarin, lignanoid, organic acid, amino acid, steroidal, The ingredients such as polysaccharide have antirheumatic, analgesia, adjust the bioactivity such as cardiovascular system, anticancer, anti-aging, radiation protection, have Very big medicinal Development volue.The raising immunity of Herba Saussureae Involueratae and its extract, anti-inflammatory, anti-spoke are described in many documents Penetrate effect." University Of Suzhou ", Gao Bo, 2001, " Antiradiation injury of Herba Saussureae Involueratae extract acts on and Mechanism Study ", it is indicated that day Mountain lanatechead saussurea herb with flower is in use for a long time as conventional medicament, and pharmacological action is clear, has the function of anti-oxidant, removing free radical well, This article passes through Integral animal experiment, Hydroxyl radical-scavenging experiment, the Giemsa staining analysis of chromosome aberration and regulating cell The RT-PCR of the P21 gene expression in period is analyzed, systematically demonstrate Herba Saussureae Involueratae extract Antiradiation injury effect and Its possible mechanism of action." Chinese radiation hygiene ", 4 phases in 2010, Guo Na etc., " Saus-surea obvallata (DC.) Edgew water extract is thin to mouse propagation The research of born of the same parents' Study On The Radioprotective " has studied Saus-surea obvallata (DC.) Edgew water extract to the Study On The Radioprotective of mouse propagation cell." Chinese medicine Pharmacology and clinic ", Huang Hua etc., " Pharmacology and Clinics of Chinese Materia Medica ", and 2002, " invigorating blood circulation analgesia and anti-inflammatory effect of saussurea involucrata frondosa oral liquid are ground Study carefully ", have studied the raising immunity and promoting blood circulation, analgesia and anti-inflammatory effect of saussurea involucrata frondosa oral liquid.
Root of kirilow rhodiola has effects that QI invigorating, promoting blood circulation, changes and become silted up." Chinese Clinical pharmacology and acology ", Jiang Ailing etc. are " red The progress of red-spotted stonecrop glycosides pharmacological action ", 2015, it is indicated that rhodioside is the main active of China's tradition Tibetan medicine root of kirilow rhodiola One of, medical value is very high, and this article makees the protection of skeletal system, cardiovascular system, nervous system, liver from rhodioside With to the therapeutic effect of diabetic nephropathy, anti-inflammatory, improving several directions such as immunity and summarize its pharmacological action, be Medical treatment research and the clinical application of related disease provide theoretical foundation." practical hospital clinical magazine ", Xiao Yu etc., " red scape Its glycosides pharmacological research progress ", 2006,3 (3): 98-99, root of kirilow rhodiola pharmacological action have antifatigue, anti-aging, immunological regulation, clearly Except free radical, anti-hepatic fibrosis, anti-renal damage improves protection cardio cerebral function, antitumor, anti-radiation to wait many-sided pharmacological action, It has a extensive future.
CN101264120B discloses a kind of barrenwort flavonol glycosides medicine materical crude slice, is made of Herba Epimedii effective part extract, Icariin content is 10-20%wt, and icariside I I content is 2-5%wt, general flavone content 40-60%wt, total Huang Ketone include icariside I I, newly determine towards leaves of pulse plants glycosides 1, towards the leaves of pulse plants K, icariin, towards leaves of pulse plants glycosides I, towards leaves of pulse plants glycosides third, towards leaves of pulse plants glycosides A, newly towards the leaves of pulse plants Glycosides 2 and Hyperoside, the preparation method of barrenwort flavonol glycosides medicine materical crude slice include: to weigh smashed korean epimedium herb overground part Point, with ethanol water refluxing extraction；Extracting solution filtering is concentrated under reduced pressure after merging, then is diluted with ethanol water, after standing from The heart, upper Diaion-HP20 resin column are collected 60-70%v ethanol water and are eluted liquid, decompression with ethanol water gradient elution It is concentrated into paste, crushed after being dried obtains barrenwort flavonol glycosides medicine materical crude slice.
CN101843629A discloses the new application of icariin and the epimedium flavone containing icariin, and in particular to The extract preparation for treat, prevent, mitigate and/or alleviate disease related with nervous system myelin disease damage and/or Purposes in the drug of illness, or in preparation for mitigating the purposes in myelinoclasis and/or the drug for promoting myelin reparation.
CN104711300A discloses a kind of preparation method of icariine, and this method includes by Shorthorned Epimedium P.E in fruit Carry out enzyme digestion reaction under the action of glue enzyme, obtain icariine, it is preferable that the pectase include polygalacturonase and Arabinofuranosidase.
CN101933550A discloses the electuary containing Herba Epimedii, and the electuary is by Herba Epimedii, ground coffee, cordyceps sinensis or pupa Cordyceps sinensis is constituted, and the weight percent of each material is as follows: Herba Epimedii and the sum of cordyceps sinensis or Cordyceps militaris account for 2-99%, ground coffee 1- 98% or Herba Epimedii 1-95%, cordyceps sinensis or Cordyceps militaris 1-96%, ground coffee 1-98%.
CN104171175A discloses a kind of Herba Epimedii tea, is prepared in following manner: a, acquisition are selected: annual Spring acquires Epimedium brevicornum leaves, chooses sundries and old leaf；B, finish: 160 DEG C~200 DEG C water-removing 1-3 minutes；C, it rubs: will Epimedium brevicornum leaves after water-removing are rubbed, and are rubbed at item；D, it fried dry: will rub to Epimedium brevicornum leaves in stripes at 130 DEG C -90 DEG C Carry out fried dry, first through 130 DEG C of fryings of high temperature, fry to it is half-dried when gradually descend temperature regulating to 110 DEG C, gradually descend temperature adjustment again through frying Degree is to 90 DEG C, until fried dry.
CN101607976A discloses a kind of preparation method of icariin, takes epimedium herb, with the methanol of 30-90% Or alcohol reflux extracts, recycling design is condensed into medicinal extract；Successively extracted repeatedly with petroleum ether, chloroform, ethyl acetate and n-butanol To close colourless, recycling n-butanol obtains coarse extract to dry；Then silica gel column chromatography separation and polyamide column chromatography point are successively carried out From to get icariin highly finished product, purity >=98%.
CN104688795A discloses a kind of method for preparing general flavone by Herba Epimedii, method includes the following steps: a. Herba Epimedii cured leaf is extracted by ethanol solution first, obtains extracting solution；B. extracting solution step a obtained is with macropore tree The absorption of rouge column, then successively uses macroporous resin column described in water and ethanol elution, collects last time ethanol eluate；It c. will step The eluent that rapid b is obtained is dry, obtains the general flavone.
CN101669980B discloses a kind of method that separating icariin and general flavone are extracted from Herba Epimedii, including with Lower step: first being extracted using 20-40% ethyl alcohol as solvent counter current, and extracting solution centrifuge separation, separating liquid is respectively with microfiltration membranes except heavy Filtering and Ultra filtration membrane, then with nanofiltration membrane treatment, obtain trapped fluid；This trapped fluid is dry, and 60 DEG C of drying temperature is hereinafter, dry Object moisture content≤5.0%, dried object are epimedium active constituent enriched substance, and wherein Icariin content reaches 60% or more, excessive Sheep leaves of pulse plants general flavone content reaches 80% or more, UF membrane each unit icariin, the epimedium flavone rate of transform >=98%.
CN101899077A discloses the extracting method of icariside I in Korean epimedium leaves, and step and condition are such as Under:
A. it is 70% alcohol reflux 4 hours with the volumetric concentration for being 15 times of its quality by Korean epimedium leaves, filters out ethyl alcohol The dregs of a decoction to flow back the volumetric concentration for being 10 times of korean epimedium herb leaf quality is that the extraction 2 of 70% alcohol reflux is small by extracting solution When, ethanol extract is filtered out, filtrate twice is merged；B. filtrate is depressurized at 50 DEG C and steams solvent, obtains extract, in extract Contained moisture is not more than the 5% of its quality；C. with gained in 2 times of korean epimedium herb leaf quality of dehydrated alcohol dissolving step b Concentrated hydrochloric acid is added, korean epimedium herb leaf quality gram in the extract arrived: the volume mL of concentrated hydrochloric acid is 1:0.05~0.15, at 50 DEG C Heating water bath 15-25 hours, left at room temperature over night, filtering；D filtrate is concentrated under reduced pressure into the 20% of its volume at 50 DEG C, and court is added Precipitating is precipitated in the distilled water of 2 times of fresh Folium Epimedii quality, is deposited at 50 DEG C and is washed with the ethanol solution that volumetric concentration is 20%, Being deposited at 50 DEG C after washing is dried in vacuo, and obtains icariside I powder.
CN105998140A discloses a kind of rhodiola root composition, is grouped as by following group: root of kirilow rhodiola 1.5-6g, thorn five Add 3-27g, Radix Angelicae Sinensis 2-12g.
CN102487979B discloses a kind of Rhodiola rosea micro-powder lotus seed paste, by weight percentage constitute include lotus seeds 50-70%, Siraitia grosvenorii 5-15%, vegetable oil 10-20%, Radix Rhodiolae micropowder 7-15%；The Radix Rhodiolae micropowder is that root of kirilow rhodiola pulverizes The micro mist that partial size made of machine is 0.1-10 μm.
CN103012512A discloses a kind of isolation and purification method of rhodioloside in natural root of kirilow rhodiola, takes natural root of kirilow rhodiola It is sliced for raw material, obtains Aqueous extracts through ultrasonic water leaching, low temperature alcohol precipitating and after being concentrated under reduced pressure, then removed through solvent Removal of impurities, solvent extraction and back extraction stage obtain the rhodioloside aqueous solution of high-purity, are most concentrated under reduced pressure afterwards, dehydrated alcohol knot Crystalline substance separates and is dried to obtain rhodioloside product.
CN101804156A discloses the preparation process and antifatigue effect of compound rhodiola root extractive, by root of kirilow rhodiola 20g-30g, rhizoma polygonati 20g-30g, Schisandra chinensis 10g-20g, Herba Epimedii 10g-20g, jujube 3g-5g, what 8-10 times of coarse powder addition was measured After water impregnates 4h-6h, 100 DEG C of heating extractions, then ultrasound 1h-1.5h, temperature is 30 DEG C, power 80W, then with gauze mistake Filter, filter residue is each primary with 6 times of amounts and 4 times of amount water repetition above steps respectively again, merges filtrate three times, people is added in vacuum drying Join three alcohol type saponin(e 0.5g-1g, obtains compound rhodiola root extractive.
" Herba Epimedii Study on extraction ", Chen Luoyi etc., Chinese Journal of Modern Applied Pharmacy, 2000 (s1): 12-14 are studied excessive Sheep leaves of pulse plants extraction process, wherein selecting L9 (3~4) orthogonal test table, knot using Icarrin as Testing index Fruit shows that decocting method is close with alcohol extracting method extraction effect, and Herba Epimedii uses decocting method, and simple process is at low cost, and extraction efficiency is high, It is comparatively ideal production technology.
By the above-mentioned prior art it is found that still having number of drawbacks in the prior art, Herba Epimedii in the prior art is extracted Object is actually the mixture of many kinds of substance, wherein including flavones ingredient, more than ten leaves of pulse plants glycosides, the leaves of pulse plants time glycosides, a variety of glucosides, the wooden rouge Element, alkaloid and polysaccharide, maltol, the vertical glycosides of sand, Quercetin, icariine A, Epimedium acuminatium glycosides, anhydroicartin, isoamyl Alkene kaempferol, ginkegetin, Isoginkgetin, bilobetin, epimidin A, Hyperoside, also containing certain Volatile oil, brown eleostearic acid, stearic acid, oleic acid, the linolenic acid of amount.In these substances, effective component is irregular or even some components Drug effect is relatively low, so that it is lower not only result in integral medicinal effect, and since some inefficient ingredients will affect high-drug-effect ingredient instead The performance of energy.In addition, there exists a further problem in that, Shorthorned Epimedium P.E and gadol extract are mixed although having in the prior art The report of compound formulation is prepared, but is usually all the extract mixing by two kinds containing complicated ingredient, not only effect is poor, but also It acts synergistically also poor.This field needs a kind of efficient Herba Epimedii and gadol extract and preparation method thereof.
Summary of the invention
To solve the above-mentioned technical problems in the prior art simultaneously, the present invention provides one kind to extract sugar from Herba Epimedii Glycoside substance method, the glucosides substance has relatively high pharmacological activity compared with other ingredients in extract, special It is not kidney tonifying benefit, essence and blood and improves immunity, anti-inflammatory efficacy and antifatigue.In addition, additionally providing comprising Shorthorned Epimedium P.E With the Chinese medicine composition of gadol extract, is acted synergistically by the compounding of the two, can be improved comprehensive pharmacological activity.
More specifically, the present invention provides a kind of raising immunity, the Chinese medicine of anti-inflammatory, radiation protection and kidney tonifying, benefiting essence-blood Composition, the composition more particularly include specifically for the Chinese medicine composition comprising Shorthorned Epimedium P.E and gadol extract The Chinese medicine composition of Saussurea involucrata extract, Shorthorned Epimedium P.E and gadol extract, that is, provide a kind of raising immunity, anti- Scorching, radiation protection and kidney tonifying, the Herba Epimedii rhodiola root composition of benefiting essence-blood or saussurea involucrata Herba Epimedii rhodiola root composition, therefore this hair Bright theme (i.e. title) also can for it is a kind of raising immunity, anti-inflammatory, antifatigue and kidney tonifying, benefiting essence-blood Herba Epimedii root of kirilow rhodiola Chinese medicine composition and preparation method thereof or a kind of raising immunity, anti-inflammatory, antifatigue and kidney tonifying, the saussurea involucrata of benefiting essence-blood are excessive Sheep leaves of pulse plants root of kirilow rhodiola Chinese medicine composition and preparation method thereof.For this purpose, the present invention provides following technical schemes.For this purpose, the present invention mentions Following technical scheme is supplied.
In one aspect of the invention, one kind is provided from plant extract glycoside substance method, and this method uses multistage column Chromatography.
Preferably, wherein the plant is Herba Epimedii.
Preferably, in the multistage column chromatography for separation, first order column chromatography uses reverse phase silica gel column.
Most preferably, the extraction and isolated glucosides substance are lower formula (I) compound represented:
Studies have shown that the glucoside compound has compared with general flavone compound such as icariine A and B Wider bioactivity, as anti-osteoporosis disease, antifatigue, androgen, anti-inflammatory and anticancer and kidney tonifying, benefiting essence-blood act on. Therefore, the application is using the compound as Objective extraction compound.
" influence of the barren wort total chromocor to rats with deficiency of kidney-Yang hypothalamus-pituitary-adrenal axis calmodulin gene expression ", Jiang Shujun etc., Chinese applied physiology magazine, 2004；Syndrome of deficiency of kidney yang pathological change and Herba Epimedii are inquired into from molecular level in 20 (4) The effect and its mechanism of general flavone provide theoretical foundation to develop the novel clinical use of drug.Including the document very More documents all report kidney tonifying including above-mentioned epimedium flavone compound (being also glucoside compound), benefiting essence-blood effect, but It is to be substantially to be studied as general flavone (i.e. master mix).
Jilin Agriculture University's master thesis, " korean epimedium herb chemical constitution study ", Wang Bin, 20070601, and north Capital university of chemical technology master thesis, " rock salt in Sichuan chemical constitution study and bacteriostatic activity research ", Song Lina, 20090601, all report numerous chromocor compounds (i.e. glucosides chemical combination including above-mentioned formula (I) and formula (II) compound Object) have effects that kidney tonifying, benefiting essence-blood, specifically, the icariin can be obviously promoted rat interstitial cell TS basal secretion, This provides experimental basis for the treatment for the male sterility that Herba Epimedii is used for TS low SI.Herba Epimedii is to glucocorticoid institute " yang deficiency syndrome " is caused to improve significantly, indices restore very fast.But above-mentioned document do not specifically note it is above two specific Compound has effects that more preferably.
" barrenwort chemical component and pharmacological research progress ", Xie Juanping, Strait Pharmaceutical Journal, 2006 volume 18 the 5th Phase has reported numerous chromocor compounds including above-mentioned formula (I) and formula (II) compound beneficial to vital essence, heavily fortified point from pharmacology angle Muscles and bones mends the effect of waist kidney, heart tonifying power.Equally, also undisclosed above two compound has more preferably kidney tonifying, benefiting essence-blood function Effect.
The present inventor has found above-mentioned formula (I) and formula (II) compound relative to other chemical combination in extract by screening For object, have effects that more preferably, as target compound, specific technique to be taken to extract, to prepare aforementioned group Close object.
For the present invention, particularly preferably, formula (I) compound represented can be made by following method, the party Method the following steps are included:
(1) Herba Epimedii herbal medicine powder is extracted (preferably with MeOH (preferably 3 × 6L, each soaking time are about 20h) at room temperature 1.18kg), combined extract is evaporated under reduced pressure to obtain brown residue (162.5g) i.e. methanol extract liquid；
(2) reverse phase silica gel column (such as RP-18 silicagel column) is utilized, uses MeOH-H first2O mixed solution is to the remnants Object carries out gradient elution, and to carry out pillar layer separation, preferred elution step is 0:1 (1L), 1:9 (1L), 2:8 (1L), 3:7 (1L), 4:6 (1L), 5:5 (2L), 6:4 (2L), 7:3 (1L), 1:0 (2L) then use acetone (2L), successively obtain 10 and wash De- component is denoted as elution component A-J；
(3) elution fraction B (such as 6.5g) is subjected to chromatographic isolation on silicagel column (preferably 60cm × 5cm), used CHCl3-MeOH-H2O mixed solution carries out gradient elution, and (gradient is preferably 20:1:0 (1L), 10:1:0 (2L), 30:8:1 (2L) and 15:6:1 (2.5L)), 4 elution components are obtained, elution fraction B1-B4 is denoted as；
(4) elution fraction B2 (such as 300.2mg) is used into MeOH-H2O (1:1) is in Sephadex LH-20 (preferably 100cm × 3cm) on separate, obtain 7 time elution fractions, be denoted as elution fraction B2.1-B2.7；
(5) by elution fraction B2.6 (such as 95.0mg) by silica gel CC column (preferably 80cm × 2.5cm), eluent is EtOAc-CHCl3-MeOH-H2O (16:8:3:1), eluent is concentrated to dryness, and obtains pale yellow powder (such as 6.1mg), As compound shown in formula (I).
It is analyzed through HPLC, the purity of compound shown in formula (I) is 99.7% in extract.Through1HNMR and mass spectral analysis, It is consistent with the known compound through characterizing recorded in CAS.Positive ion mode ESI-MS shows that molecular ion peak appears in m/z 667([M+Na]+) and 1311 ([2M+Na]+)。IR(KBr):3423,2940,1660,1605,1512,1450。1In HNMR, in δ (H) the A2B2 coupled system that 7.88 (d, J=8.6, H-C (2 ', 6 ')) and 7.11 (d, J=8.6, H-C (3 ', 5 ')) occur is shown The presence of para-orientating group in B ring；δ (H) 6.72 (d, J=10.0, H-C (1 "), 5.68 (d, J=10.0, H-C (2 "), The serial hydrogen atom signal that 2.14 (s, Me (4 ")) and 1.45 (s, Me (5 ")) occur shows that there are γ, γ-dimethyl chromene ring.
Experiment discovery, obtains compound shown in the formula (I) of high-purity to separation or extract is extremely difficult, wherein Leading to a major reason for being difficult to obtain high-purity compound is the interference of albumen in extraction process, in addition, some low liters Polysaccharide is also more difficult isolated impurity.The applicant obtains the formula of extreme high purity by above-mentioned 5 grades of column chromatography for separation for the first time (I) compound shown in.There is close level and matching relationship, setting selection and sequence are non-between 5 grades of separation systems Chang Guanjian.
Certainly, it will be appreciated by those of skill in the art that other elution fractions can also be used to obtain it is other valuable Substance rather than all discard.
In another aspect of this invention, a kind of composition comprising Shorthorned Epimedium P.E and gadol extract is provided, The Shorthorned Epimedium P.E is optionally the glucosides substance obtained according to the above method, preferably compound shown in (I).It can be with Saussurea involucrata extract, wolfberry fruit extract or other extracts with required pharmacological activity are added in the composition.
Preferably, the purity of the glucosides substance is greater than 99.0wt%.It is highly preferred that the purity of the glucosides substance Greater than 99.2wt%.Most preferably, the purity of the glucosides substance is greater than 99.7wt%.
The gadol extract is preferably rhodioside.Rhodioside can take the conventional method of this field to be mentioned It takes.It is further preferred that the gadol extract is modified gadol extract, most preferably, the gadol extract is logical It crosses to the modified lower formula (II) or (III) compound represented obtained of rhodioside:
Compared with general natural nucleoside substance, the immunological regulation of formula (II) or (III) compound represented to normal mouse Effect becomes apparent from, and due to the introducing of electron-withdrawing group Cl and F, enhances its ability for removing free radical, can dramatically increase mouse Special antibody secreting cell number can enhance delayed allergy intensity with 50mg/ (kgd) dosage, in addition also show The phagocytic function of mixed lymphocyte reaction (MLP) to special-shaped mouse and macrophage out, and can reduce to a certain extent white The activity of cytokine shows that the two is a kind of up-and-coming immunomodulator, that is, has and improve immunity, antifatigue Effect.
It can be synthesized by the following method with structure formula (II)~(III) substance: MCM- being added in the reaction vessel 22 molecular sieves alternatively property catalyst, is then added chloroform solvent, then by the acetyl of natural salidroside and chloro or fluoro Chlorine (i.e. acylating agent) is added in reaction dissolvent (the acetyl of natural salidroside and chloro or fluoro with the molar ratio of 1:1~1.3 Chlorine is preferably added in the form of chloroformic solution), reaction temperature is 20-30 DEG C, reaction time 10-30min, and separation and drying are It can get the compound.This method has the advantages that high income, impurity are few.It is particularly due to the shape-selective work of MCM-22 molecular sieve With the generation of side reaction can be significantly reduced.
In another aspect of this invention, the purposes of the composition is provided, human immunity function can be enhanced by being used to prepare Drug, health care product or the drink of energy.
Preferably, the drug is Chinese traditional compound medicine.Preferably, the composition is used to prepare oral capsule, electuary Deng.The health care product includes oral-cavity article such as chewable tablets or lozenge.
In a preferred embodiment of the present invention, in the Chinese traditional compound medicine, Shorthorned Epimedium P.E and root of kirilow rhodiola The weight ratio of derivative is 1:20-20:1.When Saussurea involucrata extract is added, the weight ratio of Saussurea involucrata extract and Shorthorned Epimedium P.E For 1:10-1:20.The commercially available acquisition of Saussurea involucrata extract improves immunity, anti-inflammatory, antifatigue and anti-radiation effect is It is known.Such composition has effects that particularly good anti-inflammatory, strengthen immunity and/or radiation protection.
Detailed description of the invention
Fig. 1 is the compound according to the embodiment of the present application 11HNMR analysis diagram；
Fig. 2 is the suppression curve figure according to the compound of the embodiment of the present application 1 to PTP1B.
Specific embodiment
Embodiment 1: the extraction of compound shown in formula (I)
Using compound shown in following operation extraction formula (I): (1) using MeOH (3 × 6L, each soaking time at room temperature It is about 20h) Herba Epimedii herbal medicine powder (1.18kg, raw material are purchased from the Tonghua, Jilin Province city town Jin Chang) is extracted, combined extract is subtracted Pressure evaporation obtains brown residue (162.5g) i.e. methanol extract liquid；(2) RP-18 reverse phase silica gel column is utilized, uses MeOH- first H2O mixed solution carries out gradient elution to the residue, and to carry out pillar layer separation, elution step is 0:1 (1L), 1:9 (1L), 2:8 (1L), 3:7 (1L), 4:6 (1L), 5:5 (2L), 6:4 (2L), 7:3 (1L), 1:0 (2L) then use acetone (2L) successively obtains 10 elution fractions, is denoted as elution component A-J；(3) by 6.5g elution fraction B silicagel column (60cm × Chromatographic isolation is carried out on 5cm), using using CHCl3-MeOH-H2O mixed solution carries out gradient elution (gradient 20:1:0 (1L), 10:1:0 (2L), 30:8:1 (2L) and 15:6:1 (2.5L)), 4 elution components are obtained, elution fraction B1-B4 is denoted as； (4) by 300.2mg elution fraction B2 MeOH-H2O (1:1) is separated on Sephadex LH-20 (100cm × 3cm), obtains 7 A secondary elution fraction, is denoted as elution fraction B2.1-B2.7；(5) by 95.0mg elution fraction B2.6 by silica gel CC column (80cm × 2.5cm), eluent EtOAc-CHCl3-MeOH-H2O (16:8:3:1), eluent is concentrated to dryness, it is light to obtain 6.1mg Compound shown in yellow powder, as formula (I).
Embodiment 2: compound shown in formula (I) inhibits PTP 1B (PTP1B) and alpha-glucosidase Ability.Specific rejection ability is as shown in table 1 below:
Table 1: the ability of the inhibition of compound shown in formula (I) PTP 1B (PTP1B) and alpha-glucosidase
In the test, malol and acarbose are used as positive reference substance.Test method is according to standard method with micro Method carries out, specifically: this system reaction 100 μ L, PTP1B substrate p-NPP of total volume is dissolved in buffer (50mmolL- 1HEPES pH value 7.3,100mmolL-1NaCl, 0.1%BSA, 1mmolL-1DTT in), pNPP reaction is final concentration of 5mmol·L-1, extract dissolves by DMSO, takes 0.5~1 μ L of enzyme solution, diluted and mixed well by 250 μ L PBS buffer solution, taken The 160 μ L of enzyme solution diluted adds to 96 orifice plates, adds 1 μ L of DMSO (extract solvent), soft mixing, at the same by PTP1B with 35mmolL is added after 96 orifice plates incubation at room temperature 5min is added in extract-120 μ L of p-NPP, 37 DEG C are protected from light 30min, 0.5mol·L-1Sodium hydroxide terminates reaction, and microplate reader reads absorbance (A) value at wavelength 405nm, according to normalized form meter Calculate corresponding half-inhibitory concentration (IC50) value；The compound is also made to the suppression curve (referring to fig. 2) of PTP1B simultaneously.
Embodiment 3: the manufacture of the capsule comprising Shorthorned Epimedium P.E and gadol extract
The capsule manufacturing method of Shorthorned Epimedium P.E and gadol extract comprising embodiment 1, wherein the root of kirilow rhodiola Extract is compound shown in formula (II), and the weight ratio of Shorthorned Epimedium P.E and root of kirilow rhodiola derivative is 5:1, and each capsule contains 0.1g content.
Embodiment 4: anti-inflammatory effect of the capsule of testing example 3 to mice auricle swelling
Kunming mice 40 are taken, weight 150-220g, half male and half female is randomly divided into 5 groups, i.e. model group, conventional extract Capsules group (0.08gkg-1, control group, wherein Shorthorned Epimedium P.E is obtained according to the method recorded in CN101264120B, It is remaining same as Example 3), 3 Capsules group (0.08gkg of embodiment-1), daily gastric infusion 1 time, continuous 4d, in last dose 30min afterwards, every mouse right ear exterior feature two sides are applied 20 μ L of dimethylbenzene, cause mice auricle swelling model, compared with left ear, Cervical dislocation puts to death mouse after 20min, two ears is cut along auricle base line, with diameter 6mm punch respectively in left and right ear antimere Round auricle is laid, precise weighing using its difference as inflammatory swelling degree (mg), and calculates swelling inhibiting rate.
Swelling inhibiting rate=(model group swelling-administration group swelling)/model group swelling × 100%
Compared with model group mouse ear swelling degree, the compound capsule group of embodiment 3 is compared with compareing each dosage group of Capsules group The inhibiting effect of mouse ear swelling caused by paraxylene has significant difference, shows to prompt compound capsule that can mitigate caused by dimethylbenzene xylene Mice auricle swelling degree has certain anti-inflammatory effect, see the table below 2:
Table 2: the influence of compound capsule paraxylene cause mice auricle swelling
Group Dosage/gkg-1 Swelling/mg Inhibiting rate/%
Model group - 8.55±1.22 -
Embodiment 3 0.08 4.01±0.92 53.10%
Control group 0.08 5.96±1.23 30.29%
By above example clearly it can be seen from Shorthorned Epimedium P.E of the invention to PTP1B) and alpha-glucosidase With good inhibiting effect, and compared with the existing technology in extract (i.e. crude extract, the mixture of Multiple components) It compares, there is obvious better antiphlogistic effects, and then there is preferable raising immunity, and then surface has preferable antifatigue effect Fruit.
Embodiment 5: the manufacture and its benefit of the capsule comprising Shorthorned Epimedium P.E, gadol extract and Saussurea involucrata extract The test of kidney, benefiting essence-blood effect
The capsule manufacturing method of Shorthorned Epimedium P.E and gadol extract and Saussurea involucrata extract comprising embodiment 1, Wherein the gadol extract is compound shown in formula (II), and the weight ratio of Shorthorned Epimedium P.E and root of kirilow rhodiola derivative is 5: 1, the weight ratio of Saussurea involucrata extract and Shorthorned Epimedium P.E is 1:15, and each capsule contains 0.1g content.The saussurea involucrata is extracted Object derives from Xi'an Bo Lin biotech firm, and the content of volatile oil is lower than 1.2wt%, rutin, chlorogenic acid, purple in the Saussurea involucrata extract The total content of syringin and dehydro-α-curcumene is lower than 2.5wt%, has parent nucleus shown in following formula (a) in the Saussurea involucrata extract Lactone compound total content is greater than 65.0wt%:
Leydig Cells (deriving from Jiangxi Institute of Medical Sciences) is taken, is established according to Klinefelter method real Test room rat interstitial cell in vitro culture model: in each culture tube, be separately added into (1) DMEM/f12 (purchased from Sigma company, 30 μ g) (control)；(2) capsule 's content according to made from the present embodiment (30 μ g).It is passed through containing 95%O2And 5%CO2Gaseous mixture Body cultivates 4h in 35 DEG C, 90r/min tepidarium oscillator after jumping a queue, and -20 DEG C of supernatant storages are taken after centrifugation, use radioimmunoassay Survey testosterone concentration relative growth rate.After tested, using the capsule 's content according to the present embodiment relative to control group, testosterone concentration Increase 31.0%.
The embodiment sufficiently shows that composition of the invention has male sex hormone and promoting sexual gland hormone sample effect, i.e. Chinese medicine institute " kidney tonifying, benefiting essence-blood " effect of meaning.The promoting sexual gland hormone sample of this and aforementioned epimedium flavone reported in existing literature acts on one It causes, and saussurea involucrata and root of kirilow rhodiola and Herba Epimedii are used in combination to rise the secretion for promoting testosterone cooperateed with to increase.
Embodiment 6: the manufacture and its benefit of the capsule comprising Shorthorned Epimedium P.E, gadol extract and Saussurea involucrata extract The test of kidney, benefiting essence-blood effect
Embodiment 5 is repeated, difference is only that in capsule 's content that gadol extract is compound shown in formula (III).Through Test, using the capsule 's content according to the present embodiment relative to control group, testosterone concentration increases 30.9%.
The effect of comparative example 5 and 6 finds, promotes testosterone secretion is smaller with the gadol extract degree of association, mainly It is that Herba Epimedii plays a role, this is almost the same with the component and effect of existing literature report.
This written description discloses the present invention, including optimal mode using example, and also enables those skilled in the art The manufacture and use present invention.It is of the invention can patentable scope be defined by the claims, and may include this field skill Other examples that art personnel expect.If this other examples have not different from the structural elements of the literal language of claims Element, or if this other examples include the equivalent structure element with the literal language of claims without substantial differences, Then this other examples are intended within the scope of claims.In the case where not will cause inconsistent degree, by reference to It will be incorporated herein in place of all references referred to herein.

Claims (10)

1. a kind of from plant extract glycoside substance method, this method uses multistage column chromatography for separation.
2. according to the method described in claim 1, wherein the plant is Herba Epimedii.
3. method according to claim 1 or 2, wherein first order column chromatography uses in the multistage column chromatography for separation Reverse phase silica gel column.
4. a kind of Chinese medicine composition comprising Shorthorned Epimedium P.E and gadol extract, the Shorthorned Epimedium P.E is according to power The glucosides substance that benefit requires method described in any one of 1-3 to obtain.
5. composition according to claim 4, wherein the purity of the glycoside substance is greater than 99.0wt%.
6. composition according to claim 5, wherein the purity of the glycoside substance is greater than 99.2wt%.
7. composition according to claim 6, wherein the purity of the glycoside substance is greater than 99.7wt%.
8. the purposes of the composition according to any one of claim 4-7, immune function of human body can be enhanced by being used to prepare Drug, health care product or drink.
9. purposes according to claim 8, wherein the drug is Chinese traditional compound medicine.
10. purposes according to claim 9, wherein Shorthorned Epimedium P.E and root of kirilow rhodiola spread out in the Chinese traditional compound medicine The weight ratio of biology is 1:20-20:1.
CN201810856507.5A 2017-09-25 2017-09-25 A kind of kidney tonifying, benefiting essence-blood, radiation protection saussurea involucrata Herba Epimedii root of kirilow rhodiola Chinese medicine composition Pending CN108976264A (en)

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