CN102920752A - Turpinia arguta leaf ethanol reflux extract, preparation method and uses thereof - Google Patents
Turpinia arguta leaf ethanol reflux extract, preparation method and uses thereof Download PDFInfo
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Abstract
The present invention relates to uses of a turpinia ternate leaf ethanol reflux extract in preparation of drugs for neuraminidase inhibition, wherein the turpinia ternate leaf ethanol reflux extract is prepared by adding ethanol to turpinia ternate leaves and carrying out extraction. The concrete extraction method comprises: taking turpinia ternate leaves; adding 40-85% ethanol in a Turpinia arguta/ethanol (W/V) ratio of 1:5-13) to carry out reflux extraction 2-3 times, wherein each reflux extraction is performed for 1-2.5 hours; filtering; mixing the obtained filtrates; and carrying out reduced pressure concentration to obtain a thick paste or drying to obtain the turpinia ternate leaf ethanol reflux extract.
Description
Technical field
The present invention relates to a kind of turpinia Montana leaf ethanol reflux extractive and its production and use.
Background technology
Influenza virus is day by day serious at present, and influenza virus and respiratory tract disease and the systemic disease that causes are closely related, China is one of influenza country occurred frequently, not only populous, and living habit also is conducive to the relay of influenza virus, the number of times of falling ill for each person every year reaches 0.3～0.7 time not to be waited, and key population reaches 2～4 times.There is serious threat in influenza to the mankind, especially new kind influenza virus, and it not only causes other infected by microbes of Secondary cases, and can directly cause organ to destroy and the allergy causing death.But effectively do not treat at present method and the treatment means of influenza virus and disease (influenza) thereof, as carry out the inflammatory reaction treatment for clinical symptoms, cure the symptoms, not the disease.The method of present modal prevention and treatment influenza virus is to use influenza virus vaccine inoculation, non-specific anti virus herb or Western medicine oseltamivir phosphate (oseltamivir phosphate capsule)-neuraminidase inhibitor.Yet influenza virus vaccine prophylactic immunization drawback is that the inoculation crowd is selective, is not that everybody can inoculate.In addition, the influenza virus vaccine protective rate is not high, and guard time is also lacked (3～6 months).Although the non-specific anti virus herb of most lays claim to antivirus action, is not for influenza virus, and antivirus action mechanism is unclear.At many kind treatment affection of exogenous wind-cold of our former studies, the Chinese prescription of the flu that doctor trained in Western medicine is thought does not have the effect of direct resisiting influenza virus.Briefly, the Chinese prescription of not every treatment flu has the effect of clear and definite inhibition influenza virus.Western medicine " oseltamivir phosphate (oseltamivir phosphate capsule; (3R; 4R; 5S)-amino-3 (1-the third 2-ethoxyethyl acetate)-1-cyclohexene-1 carboxylic acid, ethyl ester phosphate of 4-acetamide-5-) " be neuraminidase inhibitor, infected by influenza enters cell specific inhibitory effect, but not only expensive, and have that some patients take rear appearance vomiting, feel sick, the side reactions such as insomnia, headache, stomachache, diarrhoea, dizziness, fatigue, nasal obstruction, pharyngalgia and cough.Oseltamivir phosphate capsule is that the esterase that is positioned in vivo liver and intestinal is converted into active metabolite and plays the neuraminidase effect that suppresses, and also will affect its drug effect if patient's liver and intestinal organ dysfunction are undesired.In addition, the renal insufficiency patient also wants careful usefulness.
Therefore, specific aim suppresses influenza virus and copies, suppresses influenza neuraminidase hydrolysis cell surface sialic acid, thereby cause influenza virus not to be combined with cell surface receptor and to enter in the cell, and the minimizing influenza virus will be the cause of disease that treatment influenza virus sexuality is emitted in intracellular generation.
Summary of the invention
The purpose of this invention is to provide a kind of turpinia Montana leaf ethanol reflux extractive, suppress influenza neuraminidase hydrolysis cell surface sialic acid, cause influenza virus not to be combined with cell surface receptor and to enter in the cell, copy thereby can effectively suppress influenza virus, more importantly overcome the side reaction drawback of medicine.
Another object of the present invention has provided a kind of preparation method of turpinia Montana leaf ethanol reflux extractive.
Another object of the present invention has provided the purposes of the medicine of turpinia Montana leaf ethanol reflux extractive preparation prevention and treatment influenza and complication disease thereof;
Another object of the present invention has provided the pharmaceutical composition that contains turpinia Montana leaf ethanol reflux extractive.
Turpinia Montana leaf ethanol reflux extractive of the present invention, it obtains by Turpinia pomifera(Roxb) D O. is added ethanol extraction, and concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., add 30%～90% alcohol reflux 1～3 time of 5～15 times of volumes, each reflux, extract, 1～3 hour filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Preferably, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., add 40%～85% alcohol reflux 2～3 times of 5～13 times of volumes, each reflux, extract, 1～2.5 hour filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Preferred, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., add 50%～80% alcohol reflux 2～3 times of 5～11 times of volumes, each reflux, extract, 1～2 hour filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Preferred, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., add 55%～75% alcohol reflux 2～3 times of 5～10 times of volumes, each reflux, extract, 1～1.5 hour filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Preferred, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., at every turn with 60～75% ethanol of 6～9 times of volumes as extracting solvent, reflux, extract, 2 times, each 2 hours, merge extractive liquid,, filter, be evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Preferably, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., at every turn with 65% ethanol of 7 times of volumes as extracting solvent, reflux, extract, 2 times, each 2 hours, merge extractive liquid, filtered, and was evaporated to thick paste or dry turpinia Montana leaf ethanol reflux extractive.
Preferred, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., at every turn with 70% ethanol of 8 times of volumes as extracting solvent, reflux, extract, 2 times, each 2 hours, merge extractive liquid,, filter, be evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
The present invention also provides the purposes of turpinia Montana leaf ethanol reflux extractive for the preparation of the medicine of prevention and treatment influenza and complication disease thereof.
Preferably, turpinia Montana leaf ethanol reflux extractive is used for suppressing influenza virus.
Preferably, turpinia Montana leaf ethanol reflux extractive is used for suppressing influenza virus FM1.
Preferably, turpinia Montana leaf ethanol reflux extractive is used for suppressing neuraminidase.
Preferably, its complication refers to renal failure.
Preferably, its complication refers to splenic trauma or/and injury of lung.
The present invention also provides a kind of turpinia Montana leaf ethanol reflux extractive preparation, and said preparation is take turpinia Montana leaf ethanol reflux extractive as main active.
Turpinia Montana leaf ethanol reflux extractive of the present invention can extract effective inhibition neuraminic acid enzyme component, turpinia Montana leaf ethanol reflux extractive is along with the using dosage size variation, it suppresses the ability of neuraminic acid enzymatic activity, be also corresponding the changing of height of neuraminic acid enzyme inhibition rate, and one-tenth positive correlation, turpinia Montana leaf ethanol reflux extractive can be by suppressing influenza virus surface neuraminidase, and then the inhibition influenza virus enters the cell the inside, inhibition has entered the influenza virus of cell the inside and has copied, propagation, thereby reduced the infection of influenza virus to cell, growth, and prevention and treatment influenza and complication thereof.
The specific embodiment
Turpinia Montana leaf ethanol reflux extractive of the present invention, it obtains by Turpinia pomifera(Roxb) D O. is added ethanol extraction, and concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., add 5～15 times of amount 30%～90% alcohol reflux 1～3 time, each reflux, extract, 1～3 hour filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Embodiment 1
Preparation method: get leaf of Turpinia pomifera (Roxb) D O., add 13 times of amount 45% alcohol reflux 3 times, each reflux, extract, 2.5 hours filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Embodiment 2
Preparation method: get leaf of Turpinia pomifera (Roxb) D O., add 11 times of amount 50% alcohol reflux 3 times, each reflux, extract, 2 hours filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Embodiment 3
Preparation method: get leaf of Turpinia pomifera (Roxb) D O., add 12 times of amount 70% alcohol reflux 3 times, each reflux, extract, 2 hours filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Embodiment 4
Preparation method: get leaf of Turpinia pomifera (Roxb) D O., add 9 times of amount 65% alcohol reflux 3 times, each reflux, extract, 2 hours filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Embodiment 5
Preparation method: get leaf of Turpinia pomifera (Roxb) D O., add 8 times of amount 70% alcohol reflux 2 times, each reflux, extract, 2 hours filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Embodiment 6
Preparation method: get leaf of Turpinia pomifera (Roxb) D O., add 7 times of amount 65% alcohol reflux 2 times, each reflux, extract, 2 hours filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Embodiment 7
Preparation method: get leaf of Turpinia pomifera (Roxb) D O., add 9 times of amount 68% alcohol reflux 2 times, each reflux, extract, 2 hours filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Embodiment 8
Preparation method: get leaf of Turpinia pomifera (Roxb) D O., add 5 times of amount 85% alcohol reflux 1 time, reflux, extract, 1 hour filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
Experimental data 1, extraction process optimization research
Leaf of Turpinia pomifera (Roxb) D O. mainly contains the chemical compounds such as flavonoid, organic acid, flavone compound is better with the ethanol extraction effect, therefore, we with concentration of alcohol (A), add four of amount of alcohol (B), extraction time (C), extraction time (D) as the investigation factor, each factor is established three levels, presses L9 (3
4) orthogonal table carries out orthogonal (table 1, table 2) extraction process is optimized research, investigating index is the total flavones extraction ratio.
Table 1 factor level table
Get 27 parts of leaf of Turpinia pomifera (Roxb) D O. medical materials, every group average three parts, totally nine groups, by table 1L9 (3
4) the orthogonal test table design experiment.The results are shown in Table 2.
Table 2L9 (3
4) orthogonal test table
Variance analysis
Orthogonal experiments is carried out variance analysis, the results are shown in Table 3.
The variance analysis of table 3 take the total flavones extraction ratio as index
＊F
1-0.05(2，2)＝19.0
＊＊F
1-0.01(2，2)＝99.0
Conclusion: intuitive analysis as can be known, extraction time (C) and to add amount of alcohol (B) larger on extraction effect impact, concentration of alcohol (A) and extraction time (D) are less on the extraction effect impact.Table 3 the results of analysis of variance has further been verified the intuitive analysis conclusion, and namely factor C, B have considerable influence to extraction effect successively.According to producing practical situation, analyze in conjunction with the K value, determine that extraction conditions is: get leaf of Turpinia pomifera (Roxb) D O., add 10 times of amount 70% ethanol and carry 3 times, extract 2 hours at every turn, and get final product.
Experimental data 2: turpinia Montana leaf ethanol reflux extractive is to the inhibitory action of neuraminic acid enzymatic activity
Get the turpinia Montana leaf ethanol reflux extractive that embodiment 4 preparation methoies obtain, add suitable quantity of water and make dissolving, use tiring of neuraminidase inhibitor identification kit mensuration turpinia Montana leaf ethanol reflux extractive inhibition neuraminidase (N1) and see Table 4.
(1). standard curve is prepared: a. every hole in 96 hole luciferase targets adds 70 μ l neuraminidases and detects buffer; B. every hole adds respectively 0,1,2,5,7.5,10 μ l H5N1 neuraminidases again; C. every hole adds 0～20 μ l Milli-Q water again.
(2). the preparation of sample detection: a. every hole in 96 hole luciferase targets adds 70 μ l neuraminidases and detects buffer; B. every hole adds 10 μ l H5N1 neuraminidases again; C. every hole adds 0～10 μ l turpinia Montana leaf ethanol reflux extractive sample again; D. every hole adds 0～10 μ l Milli-Q water again.
(3). detecting step:
A. vibrate the about 1min of mixing;
B.37 ℃ hatch 2min inhibitor and H5N1 neuraminidase are fully interacted, the sample of doing standard curve is also hatched together;
C. every hole adds 10 μ l neuraminidase fluorogenic substrates;
D. vibrate again the about 1min of mixing;
E.37 carry out fluoremetry after ℃ hatching 20～30min.Excitation wavelength is 360nm, and emission wavelength is 440nm.
(4). calculate sample for H5N1 neuraminic acid enzymeinhibition percentage ratio according to standard curve, and calculate turpinia Montana leaf ethanol reflux extractive for the IC50 of H5N1 neuraminidase after doing concentration curve.The suppression ratio IC50 that turpinia Montana leaf ethanol reflux extractive reaches neuraminidase is 9.97mg/ml.See Table 4.
Table 4. turpinia Montana leaf ethanol reflux extractive suppresses the activity of neuraminidase
Can be clear that according to above-mentioned experimental result:
(1). turpinia Montana leaf ethanol reflux extractive can extract effective inhibition neuraminic acid enzyme component;
(2). turpinia Montana leaf ethanol reflux extractive is along with the using dosage size variation, and it suppresses the ability of neuraminic acid enzymatic activity, i.e. also corresponding changing of the height of neuraminic acid enzyme inhibition rate, and become positive correlation;
(3). by above-mentioned experiment as seen, turpinia Montana leaf ethanol reflux extractive can be by suppressing influenza virus surface neuraminidase, and then suppress that influenza virus enters the cell the inside, the influenza virus that suppresses to have entered the cell the inside copies, breeds, thereby reduced infection, the growth of influenza virus to cell, and prevention and treatment influenza and complication thereof.
Medical science and study of pharmacy personnel can't not do the inhibition influenza infection, copy in advance, or under the prerequisite of the experiment of inhibition neuraminidase, learn that in advance turpinia Montana leaf ethanol reflux extractive has prevention and treats the good result that the influenza virus sexuality is emitted.
Experimental data 3: the turpinia Montana leaf ethanol reflux extractive infected by influenza infects the inhibitory action of Embryo Gallus domesticus
Get turpinia Montana leaf ethanol reflux extractive, add suitable quantity of water and make dissolving, use influenza virus A-prime Mus lung adapted strain (FM1) and (H1N1) identify that turpinia Montana leaf ethanol reflux extractive suppresses the ability that the FM1 influenza virus is copied and suppresses in Embryo Gallus domesticus.(1). FM1 influenza virus liquid is inoculated in no-special pathogen chick embryo allantois intracavity in 10 day age, and every embryo 0.2ml was hatched 72 hours for 37 ℃, observed and calculate half Embryo Gallus domesticus infective dose (EID50).(2). turpinia Montana leaf ethanol reflux extractive adopts the toxic action of Embryo Gallus domesticus, physiological saline solution is done to be inoculated in no-special pathogen chick embryo allantois intracavity in 10 day age behind the serial dilution to turpinia Montana leaf ethanol reflux extractive, every embryo 0.2ml, each concentration is inoculated 6 embryos, hatch for 37 ℃, observe Embryo Gallus domesticus growth promoter situation, the Cmax that can survive 96 hours with Embryo Gallus domesticus is as the TD of medicine.(3). the inhibitory action of turpinia Montana leaf ethanol reflux extractive infected by influenza in Embryo Gallus domesticus adopts, 0.1ml influenza virus liquid and different dilution turpinia Montana leaf ethanol reflux extractives mix, 37 ℃ act on 2 hours, be inoculated in no-special pathogen chick embryo allantoic cavity in 10 day age, every winding kind 6 embryos were hatched 72 hours for 37 ℃.The virus attack amount is 50EID50, establishes simultaneously virus control, physiological saline solution normal control, calculates turpinia Montana leaf ethanol reflux extractive to the median effective dose (ED50) of viral inhibition.
(1) the .FM1 influenza virus is calculated through the Reed-Muench method the virulence of Embryo Gallus domesticus, and its EID50 is 10
-5.07
(2). after turpinia Montana leaf ethanol reflux extractive was inoculated in Embryo Gallus domesticus, its growth promoter and Normal group were basically identical.Embryo Gallus domesticus was all survived in 96 hours.Embryo Gallus domesticus gives turpinia Montana leaf ethanol reflux extractive stock solution and has no chicken embryo death, so can think that TD0 is 9.0g/100ml.(3). turpinia Montana leaf ethanol reflux extractive inhibitory action of infected by influenza in Embryo Gallus domesticus sees Table 5.
Table 5. turpinia Montana leaf ethanol reflux extractive infected by influenza infects the inhibitory action of Embryo Gallus domesticus
Compare with the virus control group: * P＜0.05
As shown in Table 5, turpinia Montana leaf ethanol reflux extractive has significant inhibitory action (P＜0.05) at 22.5～90.0mg/ml infected by influenza, and ED50 is 22.5mg ± 1.92mg/ml, and TI is 16.34 ± 1.35.
Experimental data 4: turpinia Montana leaf ethanol reflux extractive affects the FM1 influenza virus
Get turpinia Montana leaf ethanol reflux extractive, add suitable quantity of water and make dissolving, use influenza virus A-prime Mus lung adapted strain (FM1) and (H1N1) identify that turpinia Montana leaf ethanol reflux extractive suppresses the ability of FM1 influenza virus virulence.(1) .FM1 adopts cell median infective dose (TCID50) micromethod to the toxicity test of Testis et Pentis Canis passage cell (MDCK).(2). turpinia Montana leaf ethanol reflux extractive adopts the DMEM of serum-free that turpinia Montana leaf ethanol reflux extractive is done to be inoculated in the mdck cell hole that forms monolayer behind the serial dilution to the toxicity test of mdck cell, every hole 100 μ l, each dilution factor repeats 4 holes, establishes simultaneously the normal cell contrast.Culture plate is put 37 ℃, 5%CO
2Cultivate in the incubator, observation of cell pathological changes every day (CPE), Continuous Observation 3 days, with " +～++ ++ " the record result, press the Reed-Muench method and calculate medicine median toxic concentration (TD50) and maximal non-toxic concentration (TD0).(3). turpinia Montana leaf ethanol reflux extractive suppresses the effect of FM1 influenza virus and measures: mdck cell 5 * 10
5/ ml, every hole 100 μ l, in 96 orifice plates, 37 ℃, 5%CO
2Cultivate in the incubator, suck culture fluid in the hole next day, add 100TCID50 influenza virus liquid, every hole 100 μ l suck supernatant behind 37 ℃ of absorption 1h.Wash 2 times with phosphate buffer (PBS), take the TD0 of medicine as the 1st hole, with the DMEM liquid of serum-free turpinia Montana leaf ethanol reflux extractive is made serial dilution again, add respectively in the above-mentioned cell that has infected virus, establish simultaneously virus control and Normal group, 37 ℃, 5%CO
2Cultivate in the incubator, observe the mdck cell characteristics of lesion that influenza virus produces every day, i.e. cell monolayer degeneration becomes circle etc., for three days on end, calculates 50% of medicine and suppresses pathological changes concentration (IC50) and therapeutic index (TI).The calculating of TI: TI=TD50/IC50, the TI value is larger, shows that the safety range of medicine is larger.With Kruskal-Walis and Mann-Whitney method of inspection comparative test group and the cytopathic difference of virus control group, drug dose and suppression ratio that virus infected cell is avoided cytopathy (CPE) occurs are carried out correlation analysis, judge whether amount validity response relation.
(1) the .FM1 influenza virus is calculated through the Reed-Muench method the virulence of mdck cell, and its TCID50 is 10
-4.81(2). the TD0 of turpinia Montana leaf ethanol reflux extractive mdck cell is respectively 1026.4mg ± 30.27mg/100ml.(3). after turpinia Montana leaf ethanol reflux extractive made serial dilution, the 100TCID50 influenza virus is carried out inhibition test, median effective dose IC50 and the TI value of calculating medicine are big or small, the results are shown in Table 6.
Table 6. turpinia Montana leaf ethanol reflux extractive is to the IC50 (mg/100ml) of FM1 influenza virus and TI (x ± s)
As shown in Table 6, the cytopathogenic effect of turpinia Montana leaf ethanol reflux extractive inhibition FM1 influenza virus all strengthens along with the increase of drug dose.Drug dose and medicine are shown that to the correlation analysis that the suppression ratio of CPE carries out the dosage of turpinia Montana leaf ethanol reflux extractive and medicine are to being obvious positive correlation between the CPE suppression ratio.
Experimental data 5: turpinia Montana leaf ethanol reflux extractive is to the death protection result of influenza virus infection FM1 strain in the Mice Body
Get turpinia Montana leaf ethanol reflux extractive, use influenza virus A-prime Mus lung adapted strain (FM1) and (H1N1) identify that turpinia Montana leaf ethanol reflux extractive is to the dead protective effect of influenza virus infection FM1 strain in the Mice Body.(1). influenza virus FM1 strain virus is inoculated respectively every group of 10 BALB/C mice, male and female half and half after doing 10 times of doubling dilutions.After the slight anesthesia of ether, give and different dilution viruses respectively for every group, every mice collunarium is inoculated 20 μ l.Observe 10 days dead mouse situation, calculating LD50 by the Reed-Muench method is 10
-1.36So determine that the used modeling concentration of experiment is 10LD50.(2). turpinia Montana leaf ethanol reflux extractive is to the dead protective effect of influenza virus infection FM1 strain in the Mice Body: Normal group, influenza virus FM1 strain virus matched group, turpinia Montana leaf ethanol reflux extractive 11.25mg/ml, 22.5mg/ml, 45.0mg/ml, 90.0mg/ml dosage group philosophy gavage, gavage capacity only are 0.4ml/.After 3 days, each is organized under the slight anesthesia of ether with 10 LD50 influenza virus FM1 strain collunarium infecting mouses, 20 μ l/ except Normal group.Normal group gives the normal saline with volume simultaneously.4 groups of administration are continued administration, Normal group and influenza virus FM1 strain virus control group administered physiological saline 8 days, and dosage is the same.Day by day observe animal morbidity and record death toll, observed altogether 14 days, calculate mortality rate (mortality rate=every group of death toll/every group of total mice * 100%), the results are shown in Table 7.(3). turpinia Montana leaf ethanol reflux extractive is on the impact of influenza virus infection FM1 strain lung index in the Mice Body: Normal group, influenza virus FM1 strain virus matched group, turpinia Montana leaf ethanol reflux extractive 11.25mg/ml, 22.5mg/ml, 45.0mg/ml, 90.0mg/ml dosage group philosophy gavage, gavage capacity only are 0.4ml/.After 3 days, each is organized under the slight anesthesia of ether with 1.0 LD50 influenza virus FM1 strain collunarium infecting mouses, 20 μ l/ except Normal group.Normal group gives the normal saline with volume simultaneously.4 groups of administration are continued administration, Normal group and influenza virus FM1 strain virus control group administered physiological saline 8 days, and dosage is the same.Behind viral infection, put to death mice on the 8th day, weigh, get lung and claim lung heavy, calculate lung index (lung index=lung quality/weight * 100%); In addition, get spleen and claim spleen heavy, calculate spleen index (spleen index=spleen quality/weight * 100%), the results are shown in Table 7.
Table 7. turpinia Montana leaf ethanol reflux extractive is to the death protection result of influenza virus infection FM1 strain in the Mice Body
Annotate: ※ ※ P＜0.01VS influenza virus model group
Table 8. turpinia Montana leaf ethanol reflux extractive is on the impact of spleen index and the lung index of influenza virus infection FM1 strain in the Mice Body
Annotate: ##P＜0.01VS Normal group ※ p＜0.05VS influenza virus model group
(1). as shown in Table 7, turpinia Montana leaf ethanol reflux extractive has significant protective effect (p＜0.01) at 45.0～90.0mg/ml influenza virus infected.
(2). as shown in Table 8, turpinia Montana leaf ethanol reflux extractive has significant effect (p＜0.05) at the lung index of 90.0mg/ml influenza virus infected.
Experimental data 6: turpinia Montana leaf ethanol reflux extractive is on the impact of chronic renal failure
1) SD rat, 200～240g, Shanghai west pul-Bi Kai laboratory animal responsibility company limited, the animal quality certification number: SCXK (Shanghai) 2003-0002
2) reagent and medicine adenine, adenine (content＞98% is the import packing, Chinese Shanghai, lot number 20010520), the turpinia Montana leaf ethanol reflux extractive that embodiment 4 preparation methoies obtain;
3) test method: male SD rat, about body weight 220g, feed 10 days normal growths with normal diet first after, be divided at random Normal group, administration experimental group and modeling matched group by body weight, 12 every group.Administration experimental group and modeling matched group are made chronic renal failure (CRF) model with the adenine gavage, with adenine 320mg/ (kgd) gavage, only make the about 2ml/ of suspension with distilled water, totally 20 days; Administration after 20 days, the administration experimental group is with turpinia Montana leaf ethanol reflux extractive 1.2g/ (kgd) gavage, turpinia Montana leaf ethanol reflux extractive is made suspension solution (0.2g/ml) with distilled water, each about 2ml/ only, the gradation gavage, Normal group and modeling matched group be with equal-volume water gavage, administration after 35 days with the rat etherization. indices is observed in the blood sampling of Mus arteria caudalis.
4) result:
Table 9. turpinia Montana leaf ethanol reflux extractive is on the impact of CRF Renal Function in Rats
Annotate: through the T check, administration experimental group and modeling matched group be " * " expression P＜0.05 relatively, and " * * " represents P＜0.01
This experiment is looked sidelong at after the purine gavage sets up rat CRF model with gland, modeling control rats lethargy, body weight obviously reduces, serum Bun, Ser obviously raise, Hb obviously descends, Ret obviously raises, show that Renal Function in Rats is impaired, after giving the turpinia Montana leaf ethanol reflux extractive treatment. rat spirit is obviously bestirred oneself, and body weight is recovered gradually, improves creatinine clearance rate, promote the discharge of metabolite, reduce serum BUN, SCr concentration, renal function and anemia are obviously improved, and show that turpinia Montana leaf ethanol reflux extractive has preventive and therapeutic effect to kidney of rats damage due to the adenine.
Can be clear that according to above-mentioned experimental result:
(1). turpinia Montana leaf ethanol reflux extractive can extract effectively preventing chronic renal failure composition;
(2). turpinia Montana leaf ethanol reflux extractive has the effect while of suppressing the neuraminic acid enzymatic activity, the effect that also has the chronic renal failure controlled, overcome oseltamivir phosphate capsule sternly to the clear ability of creatinine and renal failure crowd's side effect, otherwise also had prevention effect.
Claims (6)
1. turpinia Montana leaf ethanol reflux extractive is for the preparation of the purposes of the medicine that suppresses neuraminidase, described turpinia Montana leaf ethanol reflux extractive obtains by leaf of Turpinia pomifera (Roxb) D O. is added ethanol extraction, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., 40%～85% alcohol reflux 2～3 times that adds 5～13 times of volumes, each reflux, extract, 1～2.5 hour filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
2. purposes as claimed in claim 1, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., add 50%～80% alcohol reflux 2～3 times of 5～11 times of volumes, each reflux, extract, 1～2 hour filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
3. purposes as claimed in claim 2, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., add 55%～75% alcohol reflux 2～3 times of 5～10 times of volumes, each reflux, extract, 1～1.5 hour filters; Merging filtrate is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
4. purposes as claimed in claim 3, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., each 60～75% ethanol with 6～9 times of volumes are as the extraction solvent, reflux, extract, 2 times, each 2 hours, merge extractive liquid, filters, and is evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
5. purposes as claimed in claim 4, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., at every turn with 65% ethanol of 7 times of volumes as extracting solvent, reflux, extract, 2 times, each 2 hours, merge extractive liquid,, filter, be evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
6. purposes as claimed in claim 4, concrete extracting method is as follows: get leaf of Turpinia pomifera (Roxb) D O., at every turn with 70% ethanol of 8 times of volumes as extracting solvent, reflux, extract, 2 times, each 2 hours, merge extractive liquid,, filter, be evaporated to thick paste or the dry turpinia Montana leaf ethanol reflux extractive that gets.
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| CN1857343A (en) * | 2006-04-11 | 2006-11-08 | 徐蕾 | Chinese medicine composition for preventing and treating viral cold and throat disease and its preparing method |
| CN1872106A (en) * | 2006-04-18 | 2006-12-06 | 徐蕾 | Application of wild basil circle leaves in treating disease of virulence cold |
| CN101011435A (en) * | 2007-02-25 | 2007-08-08 | 胡军 | Shanxiangyuan leaf extract, preparation method and uses thereof |
| CN101045072A (en) * | 2007-05-08 | 2007-10-03 | 胡军 | Extractive of turnipia arguta leaves and its pharmaceutical use |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1857343A (en) * | 2006-04-11 | 2006-11-08 | 徐蕾 | Chinese medicine composition for preventing and treating viral cold and throat disease and its preparing method |
| CN1872106A (en) * | 2006-04-18 | 2006-12-06 | 徐蕾 | Application of wild basil circle leaves in treating disease of virulence cold |
| CN101011435A (en) * | 2007-02-25 | 2007-08-08 | 胡军 | Shanxiangyuan leaf extract, preparation method and uses thereof |
| CN101249118A (en) * | 2007-02-25 | 2008-08-27 | 胡军 | Chinese medicine extract and medicine use thereof |
| CN101045072A (en) * | 2007-05-08 | 2007-10-03 | 胡军 | Extractive of turnipia arguta leaves and its pharmaceutical use |
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