CN106176862A - A kind of preparation method and its usage of Kalimeris integrifolia extractive of general flavone - Google Patents

Abstract

The invention discloses the preparation method and its usage of a kind of Kalimeris integrifolia extractive of general flavone, Kalimeris integrifolia polar solvent extracts, and after extracting solution filters, 60 DEG C of concentrating under reduced pressure are dried, it is thus achieved that crude extract；By macroporous resin column upper after crude extract water dissolution, with using ethanol solution eluting after the distilled water eluting of twice column volume again, collect the eluent of ethanol solution, in 60 DEG C of vacuum drying after recycling design, obtain Kalimeris integrifolia extractive of general flavone.Pharmacodynamics test proves that this extract has prevention or the effect for the treatment of immunologic liver injury.

Description

A kind of preparation method and its usage of Kalimeris integrifolia extractive of general flavone

One, technical field

The present invention relates to the preparation method and its usage of a kind of Kalimeris integrifolia extractive of general flavone, this extract can prevent Or treatment immunologic liver injury.

Two, background technology

Hepatic injury is a kind of pathological state that multiple hepatic disease is total, and its common cause of disease is that virus infects, such as clinic The infection of the virus such as upper hepatitis B virus (HBV), hepatitis C virus (HCV), its key character is substantial amounts of in hepatic tissue Inflammatory cell infiltration, produces immunity/inflammatory response, causes the hepatic injury based on immunoreation.After Virus entry hepatocyte, can Combine with hepatocyte DNA, immunocyte or antiviral drugs while killing virus, also can infected liver thin Born of the same parents, thus cause immunologic liver injury.Its long-term existence is the important beginning causing hepatic fibrosis, liver cirrhosis, even hepatocarcinoma to occur Reason element.Existing numerous studies show, in liver, the abnormal change of immunologic function and a large amount of of oxygen-derived free radicals produce is to cause liver group One of important mechanisms knitting damage (becomes Mu, the protective effect [J] to immunological liver injury of the Flos Lonicerae total flavones, Chinese medicine recklessly Pharmacology with clinical, 2007,23 (5): 85-87), it is seen that regulation immunologic function in viral hepatitis, especially chronic hepatitis treatment During importance.

The modern remedy measures to immunologic liver injury mainly has antiviral therapy, immune modulating treatment and general treatment (recovering liver function, anti-hepatic fibrosis etc.) etc..Conventional medicine is concentrated mainly on interferon, thymosin, Th cytokine, vaccine Deng application (Fan Jianghong, the progress [J] of chronic viral hepatitis B immunotherapy medicaments, foreign medical science pharmacy fascicle, 2002,29 (4): 222-228).Vaccine is the important measure of current prevention of liver disease, starts the whole people from the later stage eighties 20th century After HB vaccination, the hepatitis B infected rate of China is substantially reduced.But after there is hepatitis B vaccine injection in clinical practice at present The situation of antibody may not be produced, make a lot of Healthy People in vaccination in the near future, be in again in the danger infecting hepatitis not Know.Interferon, thymosin and nucleoside medicine are the antiviral drugs generally acknowledged at present.Nucleoside medicine mainly has rummy husband Fixed, adefovir ester, Entecavir etc..Lamivudine commonly uses the medicine for the treatment of hepatitis as current clinic, and its advantage is Efficiently, conveniently, without obvious toxicity, but life-time service La Laifu is fixed, may result in virus drug resistance, and therefore it is not a kind of permissible The hepatic medicine of root.In clinic while application nucleoside medicine, the most usually associating interferon such as alpha-interferon, β-interference Element and interferon α-1b, α-2b and can stimulate T cell secretion the lymphokine such as interferon and interleukin thymosin (injection Thymosin alpha 1 etc.) treat.This therapeutic alliance can regulate patient's immunity (cellular immunization, humoral immunization) function, and promotees Enter the Scavenging activity of drug on viral.But only some patients is had preferable curative effect, and to most patient's weak curative effects and pair Effect big (Zhang Guangye, Wang Yuqun, Zhang Wei, etc. Chinese medicine immune modulating treatment chronic viral hepatitis B progress [J]. Anhui tcm clinical practice magazine, 2002,14 (6): 5l8-521), easily cause the generation of virus persister, easily recur after drug withdrawal, and The course for the treatment of length, medical expense high.Owing to the effect of drugs of current anti-hepatitis virus is unsatisfactory, therefore, effective resisting is found While virus formulation is treated, study and find effective Chinese herbal and crude drugs preparations further and carry out anti-liver injury treatment, block Liver cirrhosis and the generation of hepatocarcinoma, delay disease process, tool to be of great significance.

Kalimeris integrifolia (Kalimeris integrtifolia Turcz.Ex DC.) is Compositae (Asteraceae) Herba Kalimeridis Belong to (Kalimeris) herbaceos perennial, another name entire leaf Herba Kalimeridis, wild Flos Chrysanthemi (Hubei), Hemerocallis citrina Baroni three grass, escobita, broom Intestinum Gallus domesticus, full leaf Radix Asteris, full leaf Aster indicus, Flos Chrysanthemi Indici etc., be mainly born in hillside, border, shrubbery, roadside.It is distributed widely in China , abroad also there is distribution at northeast, western part and middle part in Siberia, Muscovite east, Mongolia and Korea.In " China's book on Chinese herbal medicine " Record, Kalimeris integrifolia bitter in the mouth, cold in nature, there is heat-clearing and toxic substances removing, cough-relieving, effect of dissipating blood stasis hemostasis.Kalimeris integrifolia supplies with its young stem and leaf Cooked food, it is possible to do excellent herbage (Yan Hong. multistage microwave amplifier Kalimeris integrifolia total flavones Composition And Process optimizes [J]. Heilungkiang medicine Science, 2014,37 (04): 43-44).Among the people treating chronic bronchitis with its herb decoction already, curative effect is fine, 1970 Year, we were in order to inquire into the pharmacological action of Kalimeris integrifolia, carried out its root, stem, leaf, the antitussive of herb and central inhibitory action Research, it is demonstrated experimentally that Kalimeris integrifolia has obvious antitussive and central inhibitory action, herb effect the strongest (Shi Leming. Kalimeris integrifolia The experimentation [J] of antitussive effect. Chinese crude drug, 1990,13 (11): 36-37；Xu Qingrong. Kalimeris integrifolia is to central nervous system Inhibitory action [J]. Chinese crude drug, 1991,14 (7): 41-43).Separately test result indicate that, Kalimeris integrifolia also has the most anti- Scorching analgesic activity (Xu Qingrong. anti-inflammatory and analgesic effect research [J] of Kalimeris integrifolia. Chinese Journal of Modern Applied Pharmacy magazine, 2002,19 (03):199-201).The research of present inventor shows, Kalimeris integrifolia contains abundant Flavonoid substances, and flavone compound There is many effects such as antibacterial, antiinflammatory, antioxidation, defying age, anti-cancer and cancer-preventing, antiviral, but have no total to Kalimeris integrifolia at present The report that flavone is medicinal, also has no the report to Kalimeris integrifolia extractive of general flavone treatment immunologic liver injury.

Three, summary of the invention

It is desirable to provide the preparation method and its usage of a kind of Kalimeris integrifolia extractive of general flavone, to realize anti-liver damage Wound treatment, blocks liver cirrhosis and the generation of hepatocarcinoma.

The preparation method of Kalimeris integrifolia extractive of general flavone of the present invention, comprises the steps:

Kalimeris integrifolia polar solvent extracts, and after extracting solution filters, 60 DEG C of concentrating under reduced pressure are dried, it is thus achieved that crude extract；Will be thick Upper macroporous resin column after extract water dissolution, with using ethanol solution eluting after the distilled water eluting of twice column volume again, collects second The eluent of alcoholic solution, in 60 DEG C of vacuum drying after recycling design, obtains Kalimeris integrifolia extractive of general flavone.

Described polar solvent is the aqueous solution of ethanol, and percent by volume is 10%-100%, preferably 40%-80%.

Described macroporous resin selected from NKA-9, HPD-100, HPD-300, HPD-500, HPD-750, HPD-826, D-101, AB-8, preferably AB-8, HPD-100, HPD-300.

During eluting, the volumetric concentration of ethanol solution used is 30%-90%, preferably 90%.

During extraction, the quality of polar solvent is 9-21 times of Kalimeris integrifolia quality.

Kalimeris integrifolia extractive of general flavone of the present invention extracts from Kalimeris integrifolia and obtains, and the content of Kalimeris integrifolia total flavones is 50～95% percentage by weight.

The present invention uses ultraviolet spectrophotometry, with apigenin as reference substance, measures Kalimeris integrifolia total at 330nm wavelength The content of Kalimeris integrifolia total flavones in flavone extract.

Chromatographic condition is: Kromasil C₁₈Chromatographic column (250mm × 4.6mm), column temperature 30 DEG C；Mobile phase A: acetonitrile；B: 0.5% formic acid-water；Gradient elution, elution program is: 0～10min 91%～91%B, 10～25min 91%～86%B, 25～54min 86%～86%B, 54～55min 86%～82%B, 55～72min 82%～82%B, 72～73min 82%～70%B, 73～88min 70%～70%B, 88～89min 70%～64%B, 89～99min 64%～64% B, 99～100min 64%～58%B, 100～110min 58%～58%B, 110～112min 58%～50%B, 112 ～120min 50%～50%B, 120～122min 50%～43%B, 122～132min 43%～43%B, 132～ 134min 43%～27%B, 134～142min 27%～27%B, 142～144min 27%～13%B, 144～ 162min 13%～13%B, 162～164min 13%～0%B, 164～165min 0%～91%B；Flow velocity 1mL/min, UV-detector, when detection wavelength is 330nm, can measure the content of apigenin.

Possibly together with kalinturflavone A, kalinturflavone in the Kalimeris integrifolia extractive of general flavone of the present invention At least one of B.

Kalimeris integrifolia of the present invention is the herb that Compositae Herba Kalimeridis belongs to herbaceos perennial Kalimeris integrifolia.

The purposes of Kalimeris integrifolia extractive of general flavone of the present invention, is to have prevention or treatment immunologic liver injury medicine in preparation Application in thing.

The invention also discloses a kind of pharmaceutical composition containing Kalimeris integrifolia extractive of general flavone, prepare including the present invention Kalimeris integrifolia extractive of general flavone and pharmaceutically acceptable carrier and/or excipient.Such as, a kind of pharmaceutical composition, for Kalimeris integrifolia extractive of general flavone and pharmaceutically acceptable carrier and/or excipient mix according to a certain percentage；Also may be used Being that Kalimeris integrifolia extractive of general flavone, other Chinese medicine extract and pharmaceutically acceptable carrier and/or excipient are according to one Fixed ratio mixes.

Aforementioned pharmaceutical compositions can be configured to be available for, further according to conventional formulation method, the form that is administered, including per os or Parenteral administration forms.In being available for the form being administered, the Kalimeris integrifolia extractive of general flavone for the treatment of effective dose should be comprised.Institute Meaning " therapeutically effective amount " is at this dose, and the extract of the present invention can improve or palliate a disease symptom, maybe can suppress Or blocking-up advancing of disease.

The compositions being available for being administered can be that tablet, capsule, granule or liquid preparation are (such as oral or aseptic parenteral Solution or suspension).These compositionss being available for being administered can also be prepared as sustained-release preparation or targeting preparation as required.

The form of oral administration can be tablet, capsule, granule etc., and they can be containing conventional excipients as filled Agent lactose, sucrose, corn starch, sorbitol；Binding agent is as syrup, arabic gum, gelatin, sorbitol, hydroxypropyl methyl fiber Or polyvinylpyrrolidone；Lubricant such as magnesium stearate；Disintegrating agent such as starch, polyvinylpyrrolidone, crospovidone, crystallite fibre Tie up plain or pharmaceutically acceptable wetting agent such as sodium lauryl sulphate.

Fluid composition (such as liquid oral compositions) can be prepared as emulsion or syrup form, or they is prepared Become desciccate form, dissolve with other suitable solvent of smuggled goods before use.This type of liquid preparation can add containing conventional Add agent, such as sorbitol, syrup, methylcellulose, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose；Emulsifying agent such as lecithin, Anhydro sorbitol monoglyceride or arabic gum；Non-aqueous solvent includes edible oil, oily ester；Preservative such as P-hydroxybenzoic acid Methyl ester or propyl ester or sorbitol；And if needing to contain conventional correctives or coloring agent.

Parenteral composi (such as parenteral composition) can containing as described in reactive compound and sterile vehicle, according to The concentration used, described compound can be suspended or dissolved in this solvent, when preparing the solution of parenteral, can be by this The compositions of invention is dissolved in water for injection, and filtration sterilization, then fill and seals in suitable glass tube vial or ampoule.For increasing Add stability, said composition fill rear freezing also vacuum in glass tube vial can be removed moisture removal.Can with essentially identical method To prepare parenteral suspension, but described reactive compound is suspended in solvent rather than is dissolved in wherein, and degerming Can not be carried out by filtration.Then can be suspended in aseptic by described compound by being exposed to sterilizing in oxirane In solvent.Said composition preferably contains surfactant or wetting agent to contribute to being uniformly distributed of active component.

Pharmacological tests shows, the Kalimeris integrifolia extractive of general flavone of the present invention has prevention or treatment immunological liver damages Wound effect.

The present invention is using indexs such as mouse liver function and liver pathology inspections as criterion, results of pharmacodynamic test Show: the hepatic injury of BCG+LPS induction can cause ALT, AST, NO, T-BiL, AKP, TNF-α, INF-γ, IL-4, MDA, iNOS Level raises, and SOD, GSH-Px level reduces, and Kalimeris integrifolia extractive of general flavone can reduce immunological liver injury in mice serum Too high ALT, AST, NO, T-BiL, AKP, MDA, TNF-α, INF-γ, IL-4, iNOS level, antagonism SOD activity reduces, and rises High GSH-Px level, illustrates that Kalimeris integrifolia extractive of general flavone has the ability of enhancing body scavenging activated oxygen, points out full leaf Herba Kalimeridis extractive of general flavone have protection immunological liver injury in mice effect, its mechanism may directly remove free radical, Improve the activity of antioxidant enzyme and reduce LPO levels, and regulating immunological function, reduce NO and the product of TNF-α Lifes etc. are relevant.

The acute toxicity tests shows, Kalimeris integrifolia total flavones is through gastric infusion approach, the appetite of ICR mice, body weight No abnormality seen, LD₅₀> 3.1g/kg, show that toxicity is low, there is good drug safety.

The present invention possesses advantages below:

1, the preparation technology of the Kalimeris integrifolia extractive of general flavone that the present invention provides can effectively be enriched with, and goes the removal of impurity, its Kalimeris integrifolia general flavone content is high, and quality control can be carried out containing measuring by ultraviolet spectrophotometry and high performance liquid chromatography Fixed and high-efficiency liquid-phase fingerprint is analyzed, and the method is simple controlled.

2, Kalimeris integrifolia extractive of general flavone of the present invention has the effect significantly treating immunologic liver injury, and it act as Find first.

3, the toxicity of anxious poison testing surface Kalimeris integrifolia extractive of general flavone is low, has good drug safety.

4, the method for preparing extractive that the present invention provides is simple, quick, favorable reproducibility；Selected solvent is cheap, nontoxic.

Just because of above advantage, absolutely prove that this technique is suitable for industrialized production, there is the biggest practicality.

Four, accompanying drawing explanation

Fig. 1 is mouse liver tissue HE dyeing pathological section.

Five, detailed description of the invention

Embodiment 1:

Take Kalimeris integrifolia 50kg, extract with the 40vt% ethanol solution of medical material weight 16 times, 90 DEG C of circumfluence distillation 3 times, 2h every time；In 60 DEG C of concentrating under reduced pressure, 60 DEG C of vacuum drying, it is thus achieved that (Kalimeris integrifolia is total for crude extract 162.539g after extracting solution filtration The content of flavone is 26.51%).

Embodiment 2:

Take Kalimeris integrifolia 50kg, extract with the 40vt% ethanol solution of 16 times of medical material weight, 90 DEG C of circumfluence distillation 3 Secondary, each 2h；After extracting solution filters, 60 DEG C of concentrating under reduced pressure are dried, it is thus achieved that crude extract；By HPD-upper after crude extract water dissolution 300 macroporous resin column, with using 90vt% ethanol solution eluting after the distilled water eluting of twice column volume again, collect ethanol solution Eluent, in 60 DEG C of vacuum drying after recycling design, (Kalimeris integrifolia is the most yellow to obtain Kalimeris integrifolia extractive of general flavone 91.574g The content of ketone is 56.34%).

Embodiment 3:

Extract 400mg prepared by Example 1,2 method and micropowder silica gel 200mg, crosses 60 mesh sieves and fully mixes, add Appropriate magnesium stearate mixing, dry granulation.Crossing 40 mesh sieves, fill becomes capsule.

Embodiment 4:[pharmacodynamics test]

(1) animal packet, model are set up and dosage regimen

Animal is randomly divided into 6 groups by body weight, respectively normal group, model group, positive drug group (bifendate, Bifendate, Bif) and Kalimeris integrifolia total flavones group (TFKT group), TFKT group be respectively divided into again high, medium and low (600,400, 200mg·kg^-1) dosage group, often group 10, normally raise 5 days to adapt to environment.

Only, normal group gives within first day, to give all groups of mouse tail vein injection BCG 4mg/0.2mL/ in addition to normal group The normal saline of amount.Then 0.2mL/ (body weight 10g) is pressed to Bif group gastric infusion Bif 150mg kg^-1；TFKT is high, medium and low The group gastric infusion water-soluble TFKT of distillation 600,400,200mg kg^-1；Normal group, model group gavage give distilled water.Fill Stomach is administered and continues ten days, therebetween normal water feed, after last is administered, organizes equal tail vein injection LPS (10 μ g/ in addition to normal group 0.2mL/ is only) excite hepatic injury, fasting 16 hours after administration, then pluck eyeball and take blood, at cervical dislocation, win liver the most immediately Dirty tissue.

(2) index determining

Whole blood standing 1500r min after 45 minutes^-1Take serum after centrifugal 10 minutes, measure serum by test kit description NO, ALT, AST, T-BiL, AKP content.

Take the same site tissue of mouse liver, precise (about 0.3g), make the liver homogenate of 10% with ice normal saline Liquid, by specification measures liver homogenate MDA, SOD, iNOS, GSH-Px assay.

Take serum, use ELISA method detection TNF-α, INF-γ, IL-4 the most respectively to TNF-α, INF-γ and IL-4 monoclonal antibody The coated 96 every holes of hole PVC board add standard dilutions or sample to be checked 100 μ L, and set blank and standard curve hole, Each reagent dilutions concentration and step are carried out by test kit operating instruction, i.e. all measure at 450nm by microplate reader after terminating reaction Corresponding absorbance.

(3) liver histological inspection

Take the hepatic tissue of about 0.5cm × 1cm × 1cm from each group of Mouse Liver lobus dexter same area, 10% formaldehyde is fixed, and HE contaminates Color, by om observation liver tissue injury degree, and carries out micropathological damage grading: 0 grade, has no that obvious pathology damages Wound or liver have slight degeneration, and hepatocyte is without necrosis；1 grade, hepatic tissue cloudy swelling is with the spotty necrosis being dispersed in, and portal area is the most scorching Sexual cell infiltrates, and necrosis accounts for lobules of liver ＜ 1/4；2 grades, hepatic tissue cell cloudy swelling has spotty necrosis and collections necrosis to account for lobules of liver 1/4～1/3, hepatocyte interstitial is the most congested, and there is more inflammatory cell infiltration portal area；3 grades, hepatocyte cloudy swelling, focal necrosis, 1/3 ＜ necrosis accounts for lobules of liver≤1/2, and hepatocyte interstitial is the most congested, and portal area and surrounding have massive inflammatory cells infiltrated；4 grades, Hepatocyte is muddy, swelling, and part of hepatocytes ruptures, and lamellar is downright bad, and necrosis accounts for lobules of liver 1/2, portal area and surrounding a large amount of inflammation Sexual cell infiltrates.

(4) statistical method

Statistic software SPSS 21.0 is used to use One-way ANOVA to carry out statistical analysis experiment the data obtained, knot Fruit withRepresent.

(5) result

1, the TFKT impact on ALT, AST

Comparing with normal group, model group mice serum ALT, AST level is significantly raised；Compared with model group, each dose of TFKT Amount group all can reduce ALT level, but only TFKT high dose group has reduction effect for AST level value, the results are shown in Table 1.

Table 1 TFKT on the impact of ALT, AST (N=10)

“^##" represent and compare P < 0.01 with normal group；“^**" represent P < 0.01 compared with model group, "^*" represent and model group phase Ratio P < 0.05, lower same.

2, TFKT is on serum NO, the impact of T-BiL, AKP

Comparing with normal group, model group mice serum NO, T-BiL, AKP level is significantly raised；Compared with model group, TFKT Each dosage group all has NO, T-BiL, AKP level that reduction mice serum is too high, but for T-BiL level value, only TFKT Middle and high dosage group has reduction effect, and low dose group, without obvious effect, the results are shown in Table 2.

Table 2 TFKT on the impact of NO, T-BiL, AKP (N=10)

3, TFKT is on the impact of MDA, SOD, iNOS, GSH-Px level in liver homogenate

Comparing with normal group, model group mouse liver homogenate MDA, iNOS level is significantly raised, and SOD, GSH-Px level shows Write and reduce；Compared with model group, TFKT each dosage group all can reduce MDA, iNOS level that mouse liver homogenate is too high, TFKT Each dosage group all a certain degree of lifting SOD levels of energy, but dosage group the highest, middle can raise GSH-Px level, low dose group It is affected without significance, the results are shown in Table 3.

Table 3 TFKT on the impact of MDA, SOD, iNOS, GSH-Px (N=10)

4, TFKT is on TNF-α, the impact of INF-γ, IL-4

Use the TNF-α in ELISA method detection serum, INF-γ, IL-4, find that any dosage group in TFKT group is the most right TNF-α, the level of INF-γ, IL-4 have significant impact, the results are shown in Table 4.

Table 4 TFKT on TNF-α, INF-γ, IL-4 impact (N=10)

5, the TFKT impact on mouse liver changes in histopathology

HE coloration result display normal group Mouse Liver leaflet structure is clear, and centered by central vein, liver cell plate is radiation Shape arranges, and endochylema contaminates deeply, and portal area is without inflammatory cell infiltration；Model group mice HE stained finds that around central vein, liver is thin Born of the same parents' swelling and degeneration, part of hepatocytes ruptures, and interstitial is the most congested, and portal area and surrounding have massive inflammatory cells infiltrated, large area Lamellar is downright bad；The dosage group high, middle of TFKT all can substantially alleviate hepatic necrosis scope and degree, and inflammatory cell infiltration degree is also Relatively model group is light, but low dose group is to hepatocellular injury protection DeGrain, sees Fig. 1.

Embodiment 5:[acute toxicity test]

After this laboratory observation mice single gavages Kalimeris integrifolia extractive of general flavone, owing to absorbing produced toxic reaction And death condition.Result shows that the Kalimeris integrifolia extractive of general flavone 3.1g/kg that the present invention provides (is equivalent to crude drug amount 31g/ Kg) after gastric infusion, in 14d mice all none is dead, and the diet of mice, activity, fur color and luster, feces, mouth, eye, nose divide Secretions etc. are without exception, put to death mice after 14d, and the internal organs such as its heart of perusal, liver, lung, kidney, stomach, intestinal do not have ANOMALOUS VARIATIONS. Therefore the LD of Kalimeris integrifolia total flavones gastric infusion₅₀>3.1g/kg.

1, experiment material

1.1 animal

ICR mice, cleaning grade, 18-22g, male and female dual-purpose.

1.2 test medicine

(preparing by embodiment 1,2 process, content is big for the Kalimeris integrifolia extractive of general flavone extracted from Kalimeris integrifolia In 50%)

2, experimental technique

ICR mice 20, body weight 18～22g, male and female half and half, before experiment, water 12h is can't help in fasting.Administration group mice during experiment Every with the maximum of test medicine can gavage concentration (extract 0.13g/mL crude drug amount 1.3g/mL) and the maximum of mice can gavage Capacity (0.4mL/10g) once gives extract 5.2g/kg (being equivalent to crude drug amount 52g/kg), and control group mice gives equal-volume 0.5% CMC-Na solution.Dosage has reached maximum volume.Room temperature 25 ± 2 DEG C during experiment, the drink of mice in observation 14d Food, activity, fur, feces, mouth, eye, nasal discharge etc. react, and put to death mice with cervical dislocation, carry out anatomic observation after 14d The internal organs such as its heart, liver, lung, kidney, stomach, intestinal.

3, experimental result

During acute toxicity test, it is administered mice all none death, the diet of mice, activity, fur color and luster, excrement Just, mouth, eye, nasal discharge etc. without exception, put to death mice, the internal organs such as its heart of perusal, liver, lung, kidney, stomach, intestinal after 14d There is not ANOMALOUS VARIATIONS.

4, conclusion

The LD of Kalimeris integrifolia extractive of general flavone gastric infusion₅₀> 3.1g/kg, this result points out this extract being used for Time clinical, toxicity is little, and this explanation sample after preparation technology of the present invention processes is the most nontoxic.

Claims (9)

1. the preparation method of a Kalimeris integrifolia extractive of general flavone, it is characterised in that comprise the steps:

Kalimeris integrifolia polar solvent extracts, and after extracting solution filters, 60 DEG C of concentrating under reduced pressure are dried, it is thus achieved that crude extract；By crude extract By macroporous resin column upper after water dissolution, with using ethanol solution eluting after the distilled water eluting of twice column volume again, collect ethanol molten The eluent of liquid, in 60 DEG C of vacuum drying after recycling design, obtains Kalimeris integrifolia extractive of general flavone.

Preparation method the most according to claim 1, it is characterised in that:

Described polar solvent is the aqueous solution of ethanol, and percent by volume is 10%-100%, preferably 40%-80%.

Preparation method the most according to claim 2, it is characterised in that:

Described polar solvent is the aqueous solution of ethanol, and percent by volume is 40%-80%.

Preparation method the most according to claim 1, it is characterised in that:

Described macroporous resin is selected from NKA-9, HPD-100, HPD-300, HPD-500, HPD-750, HPD-826, D-101 or AB- 8。

Preparation method the most according to claim 4, it is characterised in that:

Described macroporous resin is selected from AB-8, HPD-100 or HPD-300.

Preparation method the most according to claim 1, it is characterised in that:

During eluting, the volumetric concentration of ethanol solution used is 30%-90%.

Preparation method the most according to claim 6, it is characterised in that:

During eluting, the volumetric concentration of ethanol solution used is 90%.

Preparation method the most according to claim 1, it is characterised in that:

During extraction, the quality of polar solvent is 9-21 times of Kalimeris integrifolia quality.

9. the purposes of the Kalimeris integrifolia extractive of general flavone of claim 1 preparation, it is characterised in that: in preparation, there is prevention Or the application in treatment immunologic liver injury medicine.