CN104374890B - A kind of Ilex kudincha C. J. Tseng extract - Google Patents
A kind of Ilex kudincha C. J. Tseng extract Download PDFInfo
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- CN104374890B CN104374890B CN201410633217.6A CN201410633217A CN104374890B CN 104374890 B CN104374890 B CN 104374890B CN 201410633217 A CN201410633217 A CN 201410633217A CN 104374890 B CN104374890 B CN 104374890B
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- ilex kudincha
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Abstract
The present invention relates to a kind of Ilex kudincha C. J. Tseng (IlexkudingchaC.J.Tseng) leaf extract having medical value and preparation method thereof, this extract is characterised by, rich in caffeoylquinic acid effective ingredient, eliminate toxic component Folium Ilicis cyanogen glycosides and amaroid, there is antioxidation, blood fat reducing, antiviral isoreactivity, can be used for manufacturing preventing and treating cardiovascular and cerebrovascular disease, hyperlipemia, the medicine of viral hepatitis or health product.
Description
Technical field
The present invention is the divisional application of No. 201110162622.0 patent applications, and the denomination of invention of No. 201110162622.0 patent applications of original bill is: a kind of Ilex kudincha C. J. Tseng extract, the applying date is: on June 16th, 2011.The invention belongs to medicinal plant extract field, relate to Ilex kudincha C. J. Tseng extract and preparation method thereof and in preparation method and application, including removing toxic component Folium Ilicis cyanogen glycosides and amaroid.
Background technology
Ilex kudincha C. J. Tseng (IlexkudingchaC.J.Tseng) is Aquifoliaceae this plant tall, is one of Folium Ilicis kind that south China is conventional, and ancient books supplement to the Herbal is in connection with in the record of this product blood fat reducing: " food of a specified duration makes us thin, removes people's fat ".Past 30 Years comes, many scholars carried out this plant studying (research and development of natural products, 2005,17 (3): 366-370；Hunan College of Traditional Chinese Medicine's journal, 2001,21 (2): 68-70).Pharmacological evaluation proves, this product water extract has antioxidation, expansion blood vessel, blood pressure lowering, blood fat reducing isoreactivity, but taste is extremely bitter, and many people are difficult to accept, and this product also shows some toxicity.This plant component is considerably complicated, that has reported just has the plurality of classes such as saponin, triterpene, sterol, volatile oil, fatty acid, aminoacid, trace element, vitamin, which kind of or which composition is the main lipid lowerers effective substance of Ilex kudincha C. J. Tseng actually, and existing open source literature not yet provides clear and definite answer.Triterpene and saponin component compare and are paid attention in recent years, have some patents of invention relevant with these compositions, such as CN101775061, CN101016328, CN1508149 etc..Phenolic constituent structure and pharmacologically active thereof about this plant are then rarely reported, and the toxic component about this plant is also indefinite.
Summary of the invention
An object of the present invention is to illustrate the adjustment blood fat active component of Ilex kudincha C. J. Tseng, discloses possible toxic component simultaneously, provides scientific basis for this plant resources of rational exploitation and utilization.Inventor is by a large amount of high flux screenings and experimentation, finally determine that total phenolics is blood fat reducing and antioxidant activity position, through chromatographic separation and purification and apply Modern spectroscopy technical measurement molecular structure, illustrate a series of caffeoylquinic acid composition, its content is up to more than 4%, thus disclosing Ilex kudincha C. J. Tseng first to regulate the effective ingredient physicochemical property of blood fat, establish solid foundation for extracting highway route design and quality control.
Caffeoylquinic acid in extract of the present invention all has having structure formula (I), wherein R1～R3In have 1～2 for coffee acyl, all the other are hydrogen, comprise six compounds (see following formula).Previous literature is not directed to the absolute configuration of dicaffeoyl quinic acid; the present inventor applies CD exciton chirality method 3 to obtaining from Ilex kudincha C. J. Tseng; 4-, 3; 5-and 4; the absolute configuration of 5-bis--O-caffeoyl guinic acid has measured; the all displays of its CD spectrum split point (-) Cotton curve, it was shown that two coffee acyls on ring are rotation relationship counterclockwise (accompanying drawing 1), so the absolute configuration of C-3 and C-5 is R.Concrete structure formula is as follows.
These caffeoylquinic acid compositions are known natural product, and they are widely present in plant kingdom, but are present in Ilex kudincha C. J. Tseng with so high content and specific ratio, one of important discovery that the present invention is unexpected and useful just.Accompanying drawing 2 is the typical HPLC chromatogram of extract of the present invention, reflects the basic situation of this total phenolics chemical composition.
In extract of the present invention, 3,5-bis--O-caffeoyl guinic acids and 4,5-DCQA are principle active component, and the two content fluctuates because of factors such as the medical material place of production, quality, extracting method, and content sum is generally 5%～95% (weight).The chemical composition of extract of the present invention is not limited in above-mentioned caffeoyl guinic acid, inventor have found that, natural caffeoylquinic acid C-7 carboxyl is all free, but because of contact alcohols solvent in extracting separation process, such as ethanol, methanol, may result in a small amount of esterification products to be formed, in total phenolic acid extract generally seldom, biological activity is also similar with before esterification for this kind of secondary metabolite.The inventors have also found that, containing trace flavones ingredient in phenolic acid, for Quercetin and glycoside thereof, such as rutin.It addition, as shown in Figure 2, extract also have some micro constitutents need to be further elucidated with.
Inventor finds simultaneously, and Ilex kudincha C. J. Tseng water soluble part taste is extremely bitter, and toxicity is obvious, wherein contains saponin component known in a large number.Particularly it was unexpectedly observed that water soluble part contains a kind of cyanogen glycoside material in a large number, its structure is identified as noval chemical compound Folium Ilicis cyanogen glycosides (ilexkudinin), and this product is white crystals, and molecular weight is 313.31, and molecular formula is C14H19NO7, fusing point 164～165 DEG C,-208°UVλmax259nm.Analyze (accompanying drawing 3) through X-ray single crystal diffraction and determine structural formula following (II):
The preparation method of Folium Ilicis cyanogen glycosides: take Ilex kudincha C. J. Tseng leaf 1kg, extracts three times by 95% reflow of alcohol, united extraction liquid, concentrating under reduced pressure, and extractum adds 5 times amount water stirring and dissolving, filters, and clear liquid adds 5% hcl acidifying to pH2, filters, filtrate 5%Na2CO3Neutralizing, concentrating under reduced pressure, upper macroporous resin column is adsorbed, and first washes with water, then washes with 30% ethanol, collects 30% ethanol elution fraction, concentrating under reduced pressure, then with high pressure preparative C18Post (10 μm, 40 × 250mm) separates, and detects wavelength 254nm, flow velocity 80ml/min, eluant is ultra-pure water, collects effluent according to going out peak situation, is merged by same unimodal flow point, concentrating under reduced pressure, place overnight, obtain white crystals 1.3g, be Folium Ilicis cyanogen glycosides.
Adopt the improvement karber's method acute LD to mouse stomach Folium Ilicis cyanogen glycosides50Measure, method: only take mice 5O, body weight 20 ± 2g, male and female half and half, be randomly divided into 5 groups, 1O/group.Increase and decrease by geometric progression, 5 dosage groups, adjacent two dose ratio 1:0.846 are set.Gavage respectively various dose Folium Ilicis cyanogen glycosides (300.0,253.8,214.7,181.6,153.6mg/kg).Observe clinical symptoms and death condition every day, continuous 1 week, it is shown that the LD that Folium Ilicis cyanogen glycosides is to Mouse oral50For 185.2mg/kg.After Folium Ilicis cyanogen glycosides is poisoning, liver and renal index raise, and main pathology is changed to liver and the obvious enlargement of kidney, renal cells swelling and degeneration, and part renal cells breaks.This test result indicate that, after Mouse oral Folium Ilicis cyanogen glycosides is poisoning, liver and kidney are subject to a degree of infringement.
Common are poison component type it is known that cyanogen glycoside is plant kingdom, this constituents through animal is oral enter digestive tract after, the hydrocyanic acid producing severe toxicity can be decomposed, cryanide ion enters after blood can rapidly and the prothetic group Fe of oxidized form cytochrome oxidase3+Ions binding so that it is lose the effect of transmission hydrogen atom electronics and activating molecules oxygen, causes histanoxia and suffocates.Poisoning often in acute attack, fast person is dead within half an hour, and symptom is generally by excited, dyspnea and proceed to slow pulse, platycoria, nystagmus, muscle spasm and convulsions immediately and dead.50mg hydrocyanic acid gets final product causing death.Therefore, the cyanogen glycosides main toxic component that to be probably in Ilex kudincha C. J. Tseng heretofore known.The discovery of this cyanogen glycosides provides clear and definite index components for limit detection harmful substance, and the safety tool ensureing Ilex kudincha C. J. Tseng extract is of great significance.In total phenolic acid extract of the present invention the actual content of Folium Ilicis cyanogen glycosides all≤0.2% (weight), health will not be worked the mischief by this residual quantity.The HPLC chromatogram (accompanying drawing 5) of contrast Ilex kudincha C. J. Tseng Leave extract and total phenolic acid extract can be seen that, the former is only second to 3,5-and 4,5-DCQA by the peak area of Folium Ilicis cyanogen glycosides, it is one of three main peaks, and the substantially not aobvious Folium Ilicis cyanogen glycosides chromatographic peak of the latter.
The preparation method that the two of the purpose of the present invention are to provide a kind of Ilex kudincha C. J. Tseng total phenolics effective site.Inventor is based on the full appreciation to this plant chemical ingredient, utilize the physicochemical property difference of other compositions such as caffeoylquinic acid and saponin, cyanogen glycosides, saccharide, pigment, triterpene, find out a unique syntheti c route, as shown in Figure 4, simple, be suitable to commercial production, it is characterised in that partly or entirely containing following sequence operating procedure:
A. take Ilex kudincha C. J. Tseng leaf, extract with water, ethanol or methanol eddy, filter, concentration, obtain extractum；
B. taking extractum, be dissolved in water, adjust pH6.5～8.5 with aqueous alkali, filter, filtrate adds sour water again and is acidified to pH1～4, filters, and precipitation washes sour water, drains, obtains sediment fraction；
C. the phenolic constituent of dissolved in filtrate is adsorbed by polyamide column, first washes the removal of impurity with water, then is washed down by phenolic constituent with the ethanol of 20%～95% (weight) concentration, and recycling design, concentrate is incorporated to sediment fraction；
D. precipitation uses alkanes organic solvent degreasing after drying；
E. precipitation organic solvent extraction, filters, concentration, obtains smart cream (free total phenolics)；
F. essence cream aqueous alkali dissolves, and filters, and concentration is dry, obtains total phenolics sodium salt.
Step a solvent for use, it is preferable that concentration is the ethanol of 20%～95% (weight)；Aqueous alkali used by step b and f is the aqueous solution of inorganic base, it is preferable that the alkali containing sodium ion, such as sodium hydroxide, sodium carbonate or sodium bicarbonate aqueous solution；Sour water used is inorganic acid aqueous solution, such as sulphuric acid or salt aqueous acid；Alkane solvents used by defat is one of petroleum ether, gasoline, hexane, hexamethylene or mixture, it is preferable that petroleum ether.Organic solvent used by step e is ethyl acetate, acetone, n-butyl alcohol, the one of ethanol or its mixed solvent, it is preferable that acetone and ethyl acetate, further preferably acetone.
Above-mentioned steps b～f alternative repetitive operation, to reach the purpose of further polishing purification.Polyamide column absorption described in step c, the repeatable purification for total phenolics, particularly it is enriched with dicaffeoylquinic acid.The Diluted Alcohol below 10% can first elute typically by water or concentration for non-phenolic constituent and single caffeoylquinic acid, and dicaffeoyl quinic acid is obtained more firm by polycaprolactam, generally uses concentration ethanol eluting more than 20%.The gained phenolic acid saponin without hemolytic and poisonous cyanogen glycosides, dicaffeoyl quinic acid content is high, and 3,5-and 4,5-DCQA content sum are up to 40%～90% (weight), and quality is easily controllable, safety is good, is adapted as injection crude drug.
The three of the object of the invention are to provide the medical usage of said extracted thing.After determining the blood lipid-lowering active fractions that total phenolics is Ilex kudincha C. J. Tseng, inventor attempts it has been carried out the more test of pesticide effectiveness, it is surprised to find, ischemic cardio cerebrovascular diseases is demonstrated the therapeutic effect of excellence by this extract, there is the obvious characteristic of high-efficiency low-toxicity, and also have performance out of the ordinary in treatment viral hepatitis and scavenging free radicals, antioxidation.Provide below the part the pharmacological results of extract of the present invention.
1. the impact on focal cerebral ischemia
Rat vein infusion extract (total phenols hydrochlorate), observe the impact of this product behavior on rats with cerebral ischemia caused by ferric chloride and infarct size, result shows, rat infusion extract 2,4, after 8mg/kg, animal behavior change and infarction size compare with saline control group and have clear improvement, and after 24h, behavior scoring reduces 46.2% (p < 0.01), 60.1% (p < 0.001), 50.8% (p < 0.001) respectively；Cerebral infarct size on average reduces 20.2% (p>0.05), 50.5% (p<0.001), 41.4% (p<0.01).
2. the impact on Rats with Microcirculation Disturbance pia mater encephali regional blood flow
Venoclysis extract (total phenols hydrochlorate), observes this product impact on rat microcirculation disturbance caused by macromolecule right rotary glycoside.It is shown that without substantially changing in sham-operation rat's pial local flow 60min；After intravenous injection macromolecule right rotary glycoside, rat brain mantle local flow significantly reduces, maximum decline 23.5 ± 6.2PU in 60min；Intravenous extract 2,4,8mg/kg group 10min after administration, namely rat brain mantle flow-reduction value is considerably less than solvent control group, more than effect lasts 60min, in 60min maximum decline respectively 18.1 ± 7.5,18.1 ± 5.5,13.4 ± 5.5PU, compare with the maximum drop-out value of solvent control group, p is respectively>0.05,<0.05,<0.001, illustrate to give extract and can alleviate intravenous macromolecule right rotary glycoside and cause the minimizing of microcirculation flow.
3. the impact on Rats with Microcirculation Disturbance blood viscosity
Previous experiment takes blood from rat aorta after terminating, and by volume 3.8% sodium citrate is added anticoagulant in whole blood by 1:9 ratio, with cone-plate type blood viscometer (7.5～150s under different cutting speeds-1) measure rat whole blood viscosity.It is shown that compare with sham operated rats, moulding group rat after intravenous injection macromolecule right rotary glycoside under different cutting speeds blood viscosity obviously higher than normal rat.Give after extract (total phenols hydrochlorate) 2mg/kg under each cutting speed rat serum viscosity and model control group without significant difference；Give after 4mg/kg at low shear rate (7.5s-1) time blood viscosity significantly lower than model control group (p < 0.05)；Give after 8mg/kg under each cutting speed rat serum viscosity all significantly lower than model control group.
4. the impact on Rat Experimental artery thrombosis
Take Wistar rat, by body weight random packet, often group 10.Control rats intravenous injection normal saline；Administration group intravenous extract (total phenols hydrochlorate) 2,4,8mg/kg, administration volume is 0.1ml/100g.Lumbar injection 20% urethane 1g/kg anesthesia during experiment, dorsal position is fixed, separate common carotid artery, stimulating electrode and temperature probe that experimental thrombus in vivo is formed instrument are hung on common carotid artery, and after administration, 10min starts to stimulate, and stimulus intensity is 2mA, close after stimulating 5min and stimulate switch, take off electrode, regulate temp controlled meter after 3min to zero-bit, record the artery thrombosis time.Result shows, rat intravenous extract 2,4, after 8mg/kg, compare with matched group, the artery thrombosis time postpones 9% (p>0.05), 63% (p<0.001), 108% (p<0.001) respectively, it was shown that the middle and high dosage of extract can substantially postpone the Rat Experimental artery thrombosis time.
5. antiplatelet aggregative activity
Take Wistar rat, by body weight random packet, often group 10.Control rats intravenous normal saline, administration group respectively intravenous extract (total phenols hydrochlorate) 2,4,8mg/kg, administration volume is 0.1ml/100g.Lumbar injection 20% urethane 1g/kg anesthesia during experiment, dorsal position is fixed, abdominal aortic blood, 3.8% sodium citrate with whole blood by 1:9 mixing anticoagulant, the centrifugal 7min of 1000rpm prepares platelet rich plasma, the centrifugal 10min of 3000rpm prepares platelet poor plasma, applying PPP autobalance platelet aggregation instrument, the normal saline group platelet aggregation percent induced by each derivant is adjusted to about 60%, derivant ADP, arachidonic acid (AA), collagen final concentration respectively 4 μm of ol/L, 2mmol/L, 20mg/ml.Observe the impact on administration group rat platelet aggregation effect.Result is in Table 1.
The impact (X ± SD, n=10) on rat platelet aggregation function of table 1 extract of the present invention
Note: compare with matched group, * p < 0.05；**p<0.01
6. acute toxicity test
Take female, male each 50 of body weight 18～22g Kunming mouse, each 5 groups it are randomly divided into respectively by sex, body weight, often group 10, intravenous injection extract (total phenols hydrochlorate) 2070,1863,1677,1509,1358mg/kg, adjacent two dosage group agent are from for 0.9, intravenous volume is 0.1ml/10g body weight, observes the toxic reaction of mice in administration one week after, dead distribution and dead animal number, and presses Bliss method calculating LD50And 95% fiducial limit.It is shown that mice about 1min occurs that autonomic activities reduces after high dose intravenous injection extract, repose, rapid breathing, conduct disorder occurs then, faints from fear, about 5～20min is dead.Not dead animal breathes after 30min and recovers normal gradually, and after 1h, behavior etc. all recover normally, all no longer occur death in later 6 days.The main organs no abnormality seens such as dead the mice gross necropsy heart, lung, liver.Female mice LD50It is 1666.65 (1556.12～1785.33) mg/kg；Male mice LD50It is 1740.76 (1614.34～1862.56) mg/kg, female animals LD50Approximate, no significant difference.
7. the therapeutical effect of pair dog myocardial ischemia and hemodynamic effects
Venoclysis extract (total phenols hydrochlorate) 2,4mg/kg, hence it is evident that improve Acute Myocardial Ischemia Tissues degree caused by ligation dog anterior descending coronary, reduce myocardial infarct size.Open chest anesthetized cardiac hemodynamics of dogs result of the test shows, each index of hemodynamics is had no significant effect by infusion compositions 1,2mg/kg；4mg/kg can reduce blood pressure, the maximum rate of change of left indoor pressure, left room acting and coronary resistance, on other hemodynamic parameters such as heart rate, left room EDP, total peripheral resistance, cardiac pumping functions without impact, show that extract passes through to alleviate cardiac afterload, coronary artery dilating and peripheral blood vessel, reduce returned blood volume and heart acting plays function of resisting myocardial ischemia.
8. the impact on hyperlipidemia rats serum lipid level
Select SD rat (180～200g) 80, male and female half and half, feed normal feedstuff 5d, survey serum total cholesterol (TC), triglyceride (TG) normal value, then five groups it are randomly divided into, i.e. model group with hyperlipemia, clofibrate group (positive controls) and the high, medium and low dosage group of extract, give high lipid food and feed.High lipid food formula: 79% normal feedstuff, 1% cholesterol, 10% yolk powder, 10% Adeps Sus domestica.Continuous high lipid food feeds 10d, takes hematometry TC value, it was demonstrated that all form hyperlipemia, and each group selection 10 simply continues to high lipid food and feeds, and gives relative medicine (NS suspension, 10ml/kg) respectively, and model group gives isometric(al) NS, and equal gavage gives.Every day 1 time, taking blood from orbital venous plexus respectively in 14d and 28d, survey TC, TG by enzyme reagent end-point method, obtained experimental data POMS-05 (random variance analysis) software statistics processes, and represents with x ± s.Result is in Table 2.
The impact (x ± s, n=10) on hyperlipemia rat TC and TG of table 2. extract of the present invention
Note: compare with high blood lipid model group, * p < 0.05, * * p < 0.01.
Data show, the high, medium and low dosage of extract all can obviously reduce hyperlipemia rat serum TC and TG level, and in dose-effect relationship, and under middle and high dosage conditions, effect is significantly stronger than positive drug control group.
9. lumbar injection and oral administration therapeutic effect to duck hepatitis B virus infection in duck body
Experiment adopts an age in days Beijing duck, through lower limb shin intravenous injection DHB, start to duck lumbar injection and 3 dosage groups of oral extract after 7 days, lumbar injection is 10,20,30mg/kg, oral administration is 30,60,120mg/kg, 1 day 2 times, be administered 10 days, observe the impact of the medicine toxicity on duck and the clear DHB DNA of Sanguis Anas domestica, and compare with acyclovir.Experiments show that: n of high dose oral group 120mg/kg, 1 day 2 times 10 days, avirulence.Intraperitoneal injection 30mg/kg group, by pairing statistics, after administration, after the 10th day and drug withdrawal, 3 days treatment clear DHBV-DNA of group Sanguis Anas domestica have highly significant decline and be remarkably decreased (P < 0.01-0.05)；Compare with respective matched group by statistics in groups, after administration after the 10th day and drug withdrawal 3 days, can highly significant with significantly decrease DHBV infected duck serum DHBV-DNA level (P < 0.01-0.05).Oral administration 60mg/kg, by pairing statistics, after drug withdrawal, 3 days treatment clear DHBV-DNA of group Sanguis Anas domestica have remarkable result (P < 0.05)；Adding up in groups, after administration, after the 10th day and drug withdrawal, 3 days treatment clear DHBV-DNA of group Sanguis Anas domestica are decreased significantly (P < 0.05).Within after oral 120mg/kg administration after the 5th day, the 10th day and drug withdrawal 3 days, by pairing statistics, the clear DHBV-DNA for the treatment of group Sanguis Anas domestica has notable and highly significant decline (P < 0.05-0.01)；Statistical disposition in groups, after administration, after the 5th day, the 10th day and drug withdrawal, 3 days treatment clear DHBV-DNA of group Sanguis Anas domestica have highly significant and are remarkably decreased (P < 0.01-0.05).Acyclovir comparison has remarkable result, and illustrative experiment is credible.Conclusion: extract intraperitoneal injection 30mg/kg；Oral administration 60-120mg/kg is effective to duck hepatitis B virus infection.
10. the impact on rat blood serum MDA content and SOD activity
Take male 20 months aged Wistar rats 40, body weight 400～500g, be randomly divided into four groups: matched group (feed normal feedstuff), the large, medium and small dosage group of extract (feed normal feedstuff add extract 60,40,20mg kg-1·d-1).Feeding 4w continuously, in experiment the 29th day, fasting 12h, Serum MDA (MDA) and superoxide dismutase (SOD) value are surveyed in femoral artery blood sampling.Using SPSS10.0 software analysis, compare and check with t between group, result is in Table 3.
The impact (x ± s, n=10) on rat blood serum MDA content and SOD activity of table 3. extract of the present invention
| Group | MDA(nmol/ml) | SOD(NU/ml) |
| Matched group | 8.92±0.56 | 181.45±7.15 |
| Extract (greatly) | 3.44±0.40* | 194.58±8.55* |
| Extract (in) | 4.26±0.24* | 190.36±8.82* |
| Extract (little) | 4.63±0.48* | 187.76±9.04** |
Compare with matched group: * p < 0.01；**p<0.05.
It is shown that extract of the present invention can reduce old rats Content of MDA, raise old rats activity of SOD in serum, compare with matched group that there were significant differences.
11. Antioxidative Activity Determination
Adopt DPPH oxidizing process; to the 1 of 3ml25ug/mL; 1-hexichol-2-bitterness acyl group (DPPH) solution adds 150ul sample (blank equivalent methanol replaces); cumulative volume 3.15ml; mixing; after placing 30 minutes in 37 DEG C, with E-722 type visible spectrophotometer at its light absorption value of 520nm place survey.Extract sample is 1mg/ml, 2mg/ml, 4mg/ml, 6mg/ml and 8mg/ml methanol solution, with 2,6 ditertiary butyl p cresol (BHT) for positive control.Sample clearance rate=[ODblank-ODsample/ODblank] × 100% to DPPH.In formula, the absorbance of ODblank:DPPH and solvent mixed liquor, the absorbance after ODsample:DPPH and example reaction.Result shows, DPPH is had extremely strong Scavenging activity, its IC by extract sample50For 2.3mg/ml, than BHT (IC504.2mg/ml) activity is higher.
The four of the object of the invention are to provide the application in preparing medicine, health product of the Ilex kudincha C. J. Tseng extract, including independent or with other medicinal license active ingredient combinations, it is equipped with suitable pharmaceutic adjuvant or the carrier pharmaceutically permitted, apply current known formulation process method, make various administered orally or parentally dosage form, such as tablet, hard capsule, drop pill, oral liquid, granule, injection, freeze-dried powder etc..
Accompanying drawing explanation
Figure 13,4-, 3,5-and 4,5-two-O-caffeoyl guinic acid CD collection of illustrative plates
The typical HPLC chromatogram of Fig. 2 extract of the present invention, chromatographic condition: chromatographic column VP-ODS, 150L × 4.6mm；Mobile phase MeOH 0.1%TFA gradient elution (time: 0min → 40min；MeOH:20% → 60%, 0.1%TFA:80% → 40%)；Wavelength 328nm；Flow velocity 1ml/min.
The X-ray single crystal diffraction structure chart of Fig. 3 Folium Ilicis cyanogen glycosides
The preparation method flow chart of Fig. 4 extract of the present invention
The HPLC of Fig. 5 extract of the present invention, crude extract and Folium Ilicis cyanogen glycosides compares, chromatographic condition: chromatographic column VP-ODS, 150L × 4.6mm；Mobile phase MeOH 0.1%TFA gradient elution (time: 0min → 40min；MeOH:20% → 60%, 0.1%TFA:80% → 40%)；Wavelength 259nm；Flow velocity 1ml/min.
Detailed description of the invention
With embodiment more specifically, method for preparing extractive of the present invention and the application in preparing all kinds of preparation thereof are illustrated below, but present invention is not limited to this.
Embodiment 1: the preparation of Ilex kudincha C. J. Tseng total phenolics
Take Ilex kudincha C. J. Tseng leaf 1kg, be ground into coarse powder, add 95% reflow of alcohol and extract 3 times, united extraction liquid, concentrating under reduced pressure, obtain alcohol extractum 280g.Extractum 1.5L purified water stirring and dissolving, dropping dilute sodium hydroxide adjusts pH value about 7, centrifugal, filters, and filtrate, with 5% hcl acidifying to pH2, stands 2 hours, filters, and precipitation washes with water to effluent about pH5, drains, must precipitate about 128g.Filtrate is added in polyamide (30～60 order) adsorption column, first washes with water, to effluent lighter and acidity be down to about pH5 time, use 80% ethanol eluting instead, collect ethanol eluent, concentrating under reduced pressure, obtain paste and be about 12g, be incorporated to sediment fraction.Precipitation is dried with petroleum ether backflow defat 3 times, filters, volatilizes, then extract 4 times with acetone reflux, united extraction liquid, recycling design, obtains total phenolics 82g.Its 3,5-and 4,5-DCQA content sum be 39% (weight), be substantially free of Folium Ilicis cyanogen glycosides, lightly seasoned not bitter, can as various oral and external preparation raw materials.Embodiment 2: the preparation of Ilex kudincha C. J. Tseng total phenolics
Take Ilex kudincha C. J. Tseng leaf 1kg, be ground into coarse powder, boiling 3 times, united extraction liquid, concentrating under reduced pressure, obtain extractum 220g.Extractum 1.5L water stirring and dissolving, adds 5% sodium carbonate and adjusts pH value about 7, centrifugal, filters, and filtrate, with 5% sulfuric acid acidation to pH2, stands 2 hours, filters, and precipitation washes with water to effluent about pH5, drains, must precipitate about 100g.Filtrate is added in polyamide (30～60 order) adsorption column, first washes with water, to effluent lighter and acidity be down to about pH5 time, use 60% ethanol eluting instead, collect ethanol eluent, concentrating under reduced pressure, obtain paste and be about 10g, be incorporated to sediment fraction.Precipitation is dried to reflux defat 3 times with hexane, filters, volatilizes, then with ethyl acetate reflux, extract, 4 times, united extraction liquid, recycling design, obtains total phenolics 85g.Its 3,5-and 4,5-DCQA content sum be about 42%, be substantially free of Folium Ilicis cyanogen glycosides, lightly seasoned not bitter, can as various oral and external preparation raw materials.
Embodiment 3: the preparation of Ilex kudincha C. J. Tseng total phenols hydrochlorate
Example 1 or 2 gained total phenolics 20g, slow gradation adds 5% Sodium hydroxide q. s, makes abundant dissolving under stirring, controls pH value of solution 7～8, filters, spray drying, obtains total phenols hydrochlorate 19g.This product good water solubility, can as the raw material of various solids and liquid preparation.
Embodiment 4: the preparation of injection Ilex kudincha C. J. Tseng total phenols hydrochlorate
Example 1 or 2 gained total phenolics 30g, adds 100ml95% ethanol and makes dissolving, admix appropriate polyamide, dry, add in polyamide column, first wash with water, then wash with 5% ethanol, then wash with 80% ethanol, collect 80% ethanol eluent, concentrating under reduced pressure, obtain paste 18g；Continuation acetone reflux extracts 4 times, united extraction liquid, recycling design, and gained paste dissolves with appropriate 5% sodium carbonate, controls pH value of solution 7～8, ultrafiltration, concentrating under reduced pressure, spray drying, obtains brownish-yellow powder 14g.Its 3,5-and 4,5-DCQA content sum be about 80% (weight), without saponin and Folium Ilicis cyanogen glycosides, good water solubility, can as injection raw material.
Embodiment 5: prepared by tablet
Take total phenolic acid extract 20g of the present invention, mix with starch 100g, dextrin 5g, add 10% starch slurry soft material, granulate with 14 order nylon screens, 60～70 DEG C of aeration-dryings, 16 mesh sieve granulate, add magnesium stearate 1.5g, sodium carboxymethyl cellulose 5g, mixing, it is pressed into 1000, coating, to obtain final product, and every containing extract 20mg.Dosage and number of times are determined according to Clinical efficacy.
Embodiment 6: prepared by capsule
Take total phenolic acid extract 20g of the present invention, mix with starch 120g, magnesium stearate 2g, directly fill into 1000 with Autocapsulefillingmachine, polishing, to obtain final product, every containing extract 20mg.Dosage and number of times are determined according to Clinical efficacy.
Embodiment 7: prepared by drop pill
Take total phenolic acid extract 12g of the present invention, put in the 32g polyethylene glycol 6000 adding heat fusing, stirring to dissolve, be transferred in reservoir, airtight and be incubated at 80～90 DEG C, regulate pill dripping machine drop quantitative valve, instilling in the liquid Paraffin of 10～15 DEG C from top to bottom, system 1000, drains the drop pill of formation and wipes liquid Paraffin altogether, being drying to obtain, every containing extract 12mg.Dosage and number of times are determined according to Clinical efficacy.
Embodiment 8: prepared by oral liquid
Taking total phenols hydrochlorate extract 20g of the present invention, mix with Mel 400g, sucrose 100g, sodium benzoate 6g and distilled water 2000ml, heating, to 85～90 DEG C, is stirred to dissolve, insulation 30min, filters, and filtrate is diluted with water to 10000ml, stirs evenly, embedding (every 10ml), sterilizing, to obtain final product.
Embodiment 9: prepared by granule
Take total phenolic acid extract 4g of the present invention, dextrin 16g, cane sugar powder 230g and appropriate amount of ethanol, mixing, cross 10 mesh sieves and make granule, dry in 60～70 DEG C, granulate, subpackage, to obtain final product, often bag weight 2.5g.
Embodiment 10: prepared by injection
Take total phenols hydrochlorate 50g of the present invention, inject and make dissolving with water in right amount, 0.02% activated carbon adding amount of preparation stirs 5～10 minutes, filters, and filtrate is diluted to about 10L, add sodium chloride adjustment infiltration and be depressed into isotonic, adjusting pH7.5～8.0, filter, embedding becomes 1000 (10ml/ props up), sterilizing, to obtain final product.It is available for intravenous administration.Dosage and number of times are determined according to Clinical efficacy.
Embodiment 11: prepared by freeze-dried powder
Total phenols hydrochlorate 50g of the present invention is taken under aseptic condition, it is placed in sterile chamber, inject and be about 900ml with water, it is stirred to dissolve, regulate pH value to 7.0～7.5, inject water to 1000ml, then add 0.02% activated carbon stirring 5～10min of amount of preparation, filter with aseptic suction funnel, ultrafiltration, filtrate is sub-packed in ampoule after the assay was approved, frozen drying, aseptic sealing by fusing and get final product, every 50mg, inject before use and make dissolving with water in right amount, with slowly intravenous drip after sodium chloride transfusion 250～500ml dilution.Dosage and number of times are determined according to Clinical efficacy.
Embodiment 12: prepared by compound capsule
Take total phenolic acid extract 20g of the present invention, mix with arasaponin 20g, starch 150g and ethanol in proper amount, cross 10 mesh sieves, dry, granulate, add magnesium stearate 2g, mixing, fill into 1000 with Autocapsulefillingmachine, polishing, to obtain final product.Every containing each 20mg of extract and arasaponin.The orally available preventing and treating for various ischemic cardio cerebrovascular diseases.Dosage and number of times are determined according to Clinical efficacy.
Claims (2)
1. there is the Folium Ilicis cyanogen glycosides application as detection Ilex kudincha C. J. Tseng extract and/or its preparation harmful substance index components of formula II below structure,
2. application according to claim 1, it is characterised in that actual content≤0.2% (weight) in total phenolic acid extract as detection Ilex kudincha C. J. Tseng extract and/or the Folium Ilicis cyanogen glycosides of its preparation harmful substance index components.
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Non-Patent Citations (4)
| Title |
|---|
| Cyanogenic glycosides and menisdaurin from Guazuma ulmifolia，Ostrya virginiana,Tiquilia plicata,and Tiquilia canescens;David S.Seigler等;《Phytochemistry》;20051231;第66卷;第1567-1580段 * |
| STRUCTURAL REVISION OF A PUTATIVE CYANOGENIC GLUCOSIDE FROM ILEX AQUIFOLIUM;ADOLF NAHRSTEDT等;《Phytochemistry》;19901231;第29卷(第12期);第3934 3936页 * |
| STRUCTURE OF A CYANOGLUCOSIDE OF LITHOSPERMUM PURPUREO-CAERULEUM;A.SOSA等;《Phytochemntry》;19771231;第16卷;第707-709页 * |
| 苦丁茶水提物的毒理学研究;许实波等;《中山大学学报(自然科学版)》;20010531;第40卷(第3期);第92-94页 * |
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