CN102696638B - Sweroside pharmaceutical preparation and application thereof in prevention of tobacco mosaic virus - Google Patents
Sweroside pharmaceutical preparation and application thereof in prevention of tobacco mosaic virus Download PDFInfo
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- CN102696638B CN102696638B CN2012101155763A CN201210115576A CN102696638B CN 102696638 B CN102696638 B CN 102696638B CN 2012101155763 A CN2012101155763 A CN 2012101155763A CN 201210115576 A CN201210115576 A CN 201210115576A CN 102696638 B CN102696638 B CN 102696638B
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Abstract
The invention discloses a sweroside pharmaceutical preparation and application thereof in prevention of a tobacco mosaic virus. The sweroside pharmaceutical preparation comprises 0.1-99% of sweroside and the balance pharmaceutic adjuvant and/excipient which is/are acceptable in pharmacology. The sweroside used in the sweroside pharmaceutical preparation can be separated and prepared from swertia cincta burk, the method is simple and the extraction rate is high. The sweroside pharmaceutical preparation has wide application prospects in prevention of the tobacco mosaic virus. The sweroside is separated and obtained from the swertia cincta burk through a modern separation technology. An experiment shows that the sweroside can significantly inhibit the tobacco mosaic virus (TMV) infection activity and serves as a novel plant virus resistance activity ingredient to be used for preventing a TMV disease, and the inhibition effect of the sweroside pharmaceutical preparation is better than that of other sold TMV resistant plant preparations (such as ningnanmycin). The sweroside pharmaceutical preparation is a pure biological agent, is harmless to people and animals, has a significant effect on TMV disease prevention and can be used in production of organic tobaccos.
Description
Technical field
The invention belongs to medicinal plants and applied technical field, be specifically related to a kind of sweroside pharmaceutical preparation, from surrounded swertia herb (
Swertia cinctaBurk) method of extraction sweroside and the application of pharmaceutical preparation in preparation prevention and control tobacco mosaic virus disease and anti-virus formulation.
Background technology
Tobacco mosaic virus (
Tobacco mosaic virus, be the pathogene of tobacco mosaic etc. TMV), belong to the Viraceae Tobamovirus of wearing armor (
Tobamovirus).The host of TMV is extensive, can infect 300 various plants of 30 sections, and is serious through crop harm at grains such as tobacco, tomatoes, and the whole world whenever is only the loss that the mosaic disease that caused by TMV causes and is just surpassed 100,000,000 dollars, causes huge economic loss to agricultural production.Yet because TMV has not complete immune metabolic system as animal of obligate parasitism and plant to the host, therefore, plant is in case just be in throughout one's life the state that is endangered by virus infections.These factors have brought very large difficulty all for the research and development of the Antiphytoviral medicament of high selectivity.In present agricultural production, the chemical pesticide control tobacco mosaic virus disease is most widely used, but the chemical control effect is limited, and easily causes residual the exceeding standard of farming, therefore in the urgent need to the development of new anti-virus formulation.
Plant has been synthesized the abundant secondary metabolite with antimicrobial acivity of type during evolution, and they can optionally resist the infringement of the pathogen that comprises TMV.Owing to be the antiviral substance of plant generation itself, the plant resource viral inhibitors is little or do not have toxicity, mechanism of action uniqueness and aboundresources to the toxicity of host own.Therefore, screening novel plant source antivirus preparation is significant.
The process that virus produces harm to plant can be divided into infecting and copies in host, the body and symptom is expressed three parts.The harm that therefore, all can alleviate the viroses of plant to the inhibition of any part wherein.Although some anti-plant virus agent can activate the host and produce system's resistance the not effect of these three parts in addition.Simultaneously, the antiviral activity of many active substances performance may be the result that concurs of one or more mechanism wherein.Inventor's early-stage Study shows that a lot of plant resource viral inhibitors are the systemic acquired resistances by activated plant, thereby strengthens plant to the resistance of virus.
Gentianaceae Swertia plant (
Swertia) mainly originate in the ground such as Yunnan, Guizhou, Sichuan, be southwest minority autonomic drug source commonly used.This platymiscium has 170 kinds, and China has 80 kinds approximately, mainly is distributed in the southwest, and only Yunnan just has 38 kinds.This genus medicinal plant has the effects such as clearing heat secreting bile, cardiac stimulant, relaxing liver and strengthening stomach, detoxifcation mostly, is used for the treatment of the diseases such as hepatitis, icteric hepatitis, liver and gall, but not yet has this platymiscium of bibliographical information to have the effect of Antiphytoviral.
Summary of the invention
The first purpose of the present invention is to provide a kind of sweroside pharmaceutical preparation; The second purpose is to provide a kind of method of extracting sweroside; The 3rd purpose is to provide the formulation of sweroside pharmaceutical preparation; The 4th purpose is to provide the application of sweroside in the sick preparation of preparation resisting tobacco mosaic virus.
The first purpose of the present invention is achieved in that described sweroside pharmaceutical preparation contains sweroside 0.1 ~ 99%, and all the other are acceptable pharmaceutic adjuvant and/or excipient on the pharmacology.
The second purpose of the present invention is achieved in that take surrounded swertia herb as raw material, and extracts the sweroside compound by the technique that comprises the following steps:
A, get the surrounded swertia herb that dries in the shade and pulverize, with 75 ~ 85% methyl alcohol extraction 2 ~ 3 times, each 1 ~ 3h;
B, merging extract, filter cleaner, merging filtrate, then reduced pressure concentration gets the medicinal extract of relative density 1.2 ~ 1.3;
The water of C, 1 ~ 3 times of volume of medicinal extract adding is stirred to and is suspension, with concentration 95 ~ 100% dichloromethane solutions extraction 2 ~ 3 times, each 15 ~ 25min;
D, combining extraction liquid are crossed the decolouring of MCI post, use afterwards the methanol-eluted fractions of concentration 70 ~ 80%, collect eluent;
E, use the macroporous resin column desugar again, and with the eluant solution of methanol/water (V/V) 0 ~ 100:100 ~ 0, collect eluent, concentrated the medicinal extract that contains 70 ~ 80% methyl alcohol;
F, upper HPLC separate, and with 70 ~ 80% methanol aqueous solution wash-outs, carry out purifying with semi-preparative column at last, obtain the sweroside compound.
The 3rd purpose of the present invention be achieved in that by prescription requirement ratio with the sweroside compound with to the nontoxic pharmaceutically acceptable auxiliaries of humans and animals and/or mixed with excipients, be mixed with aqua, tablet, granule or microemulsion.
The 4th purpose of the present invention is achieved in that the application of described sweroside pharmaceutical preparation in preparation control tobacco mosaic virus disease preparation.
The present invention use modern separation technology from surrounded swertia herb (
Swertia cincta Burk) obtain a kind of iridoid---sweroside middle the separation.Experimental results show that sweroside has the activity that significant inhibition TMV infects, can be used as the active component of novel Antiphytoviral, can be used for preparing the pharmaceutical preparation of preventing and treating tobacco mosaic virus disease, its inhibition is better than commercially available resisting tobacco mosaic disease plant medical agent, such as medicines such as Ningnanmycins, pharmaceutical preparation of the present invention is not only nontoxic, harmless to people and animals, safe and reliable, and the control tobacco mosaic virus disease is had significant effect.
Description of drawings
Fig. 1 is the content block diagram that the TAS-ELISA method detects virus.Among Fig. 1: the negative contrast of CK-, the positive contrast of CK+, positive greater than positive control, negative less than 2 times of negative controls.Ningnanmycin is the marketed drugs contrast, and sweroside-1,5,10,15 and 20 represents that respectively the concentration of sweroside is 1 μ g/ml, 5 μ g/ml, 10 μ g/ml, 15 μ g/ml and 20 μ g/ml.
Fig. 2 is effect contrast photo of the present invention.Its left figure is the compounds of this invention and control group Contrast on effect; Right figure is existing compound and the contrast of control group effect of drugs.
Embodiment
The present invention is further illustrated below in conjunction with embodiment, but never in any form the present invention is limited, and any change or improvement based on training centre of the present invention is done all belong to protection scope of the present invention.
Sweroside pharmaceutical preparation of the present invention is: contain sweroside 0.1 ~ 99%, all the other are acceptable pharmaceutic adjuvant and/or excipient on the pharmacology.
As preferred embodiment:
Contain sweroside 0.5 ~ 90%, all the other are acceptable pharmaceutic adjuvant and/or excipient on the pharmacology.
Contain sweroside 1 ~ 50%, all the other are acceptable pharmaceutic adjuvant and/or excipient on the pharmacology.
Contain sweroside 5 ~ 20%, all the other are acceptable pharmaceutic adjuvant and/or excipient on the pharmacology.
Sweroside for the preparation of above-mentioned sweroside pharmaceutical preparation of the present invention, its preparation method is: take surrounded swertia herb as raw material, and extract the sweroside compound by the technique that comprises the following steps:
A, get the surrounded swertia herb that dries in the shade and pulverize, with 75 ~ 85% methyl alcohol extraction 2 ~ 3 times, each 1 ~ 3h;
B, merging extract, filter cleaner, merging filtrate, then reduced pressure concentration gets the medicinal extract of relative density 1.2 ~ 1.3;
The water of C, 1 ~ 3 times of volume of medicinal extract adding is stirred to and is suspension, with concentration 95 ~ 100% dichloromethane solutions extraction 2 ~ 3 times, each 15 ~ 25min;
D, combining extraction liquid are crossed the decolouring of MCI post, use afterwards the methanol-eluted fractions of concentration 70 ~ 80%, collect eluent;
E, use the macroporous resin column desugar again, and with the eluant solution of methanol/water (V/V) 0 ~ 100:100 ~ 0, collect eluent, concentrated the medicinal extract that contains 70 ~ 80% methyl alcohol;
F, upper HPLC separate, and with 70 ~ 80% methanol aqueous solution wash-outs, carry out purifying with semi-preparative column at last, obtain the sweroside compound.
As preferred embodiment:
Described macroporous resin column is any one among D900, D130, D101, D201 or the D301.
Described semi-preparative column is any one in the uncommon semi-preparative column of Ai Jieer semi-preparative column, HPLC semi-preparative column or Japanese aestheticism.Described Ai Jieer semi-preparative column carries out purifying, adopts methanol/water solution to carry out wash-out, obtains the sweroside compound.
The formulation of sweroside pharmaceutical preparation of the present invention is: with the sweroside compound and to the nontoxic pharmaceutically acceptable auxiliaries of humans and animals and/or mixed with excipients, be mixed with aqua, tablet, granule or microemulsion by prescription requirement ratio.
Described pharmaceutical carrier or excipient are one or more solids, semisolid and liquid diluent, filler and pharmaceutical preparation assistant agent.The method that adopts pharmacy and field of food to generally acknowledge is prepared into various formulations, such as liquid preparation (injection, supensoid agent, emulsion, solution, syrup etc.), solid pharmaceutical preparation (tablet, capsule, granule, electuary etc.), spray, aerosol etc.Auxiliary substance, stabilizing agent be can contain in the above-mentioned preparation, humectant and other additives commonly used helped, such as starch, albumen powder, pectin, sucrose, dextrin, dolomol, mannitol, neopelex etc.Above-mentioned preparation can be made according to the preparation technology of various preparation routines.
The application of described sweroside pharmaceutical preparation in preparation control tobacco mosaic virus disease preparation, wherein the sweroside solution concentration is 1 ~ 20 μ g/ml.
Embodiment 1
Get the surrounded swertia herb that dries in the shade and pulverize, extract 2 times with 75% methyl alcohol, each 1h; Merge extract, filter cleaner, merging filtrate, then reduced pressure concentration gets the medicinal extract of relative density 1.2 ~ 1.3; The water of 1 times of volume of medicinal extract adding is stirred to and is suspension, with concentration 98% dichloromethane solution extraction 2 times, and each 15min; Combining extraction liquid is crossed the MCI decolouring, uses afterwards the methanol-eluted fractions of concentration 75%, collects eluent; Use again the desugar of D900 macroporous resin column, with the eluant solution of methanol/water (V/V) 10:90, collect eluent; Concentrate to get medicinal extract, upper HPLC separates, the methanol aqueous solution wash-out with 70%, and last Ai Jieer semi-preparative column carries out purifying, and the methanol aqueous solution wash-out with 70% obtains the sweroside compound.
Embodiment 2
Get the surrounded swertia herb that dries in the shade and pulverize, extract 3 times with 85% methyl alcohol, each 3h; Merge extract, filter cleaner, merging filtrate, then reduced pressure concentration gets the medicinal extract of relative density 1.2 ~ 1.3; The water of 3 times of volumes of medicinal extract adding is stirred to and is suspension, with concentration 95% dichloromethane solution extraction 3 times, and each 25min; Combining extraction liquid is crossed the decolouring of MCI post, uses afterwards the methanol-eluted fractions of concentration 70%, collects eluent; Use again the desugar of D130 macroporous resin column, with the eluant solution of methanol/water (V/V) 30:70, collect eluent; Concentrate to get medicinal extract, upper HPLC separates, 80% methanol aqueous solution wash-out, and last HPLC semi-preparative column carries out purifying, and the methanol aqueous solution wash-out with 80% obtains the sweroside compound.
Embodiment 3
Get the surrounded swertia herb that dries in the shade and pulverize, extract 3 times with 80% methyl alcohol, each 2h; Merge extract, filter cleaner, merging filtrate, then reduced pressure concentration gets the medicinal extract of relative density 1.2 ~ 1.3; The water of 2 times of volumes of medicinal extract adding is stirred to and is suspension, with concentration 100% dichloromethane solution extraction 3 times, and each 20min; Combining extraction liquid is crossed the decolouring of MCI post, uses afterwards the methanol-eluted fractions of concentration 80%, collects eluent; Use again the desugar of D101 macroporous resin column, with the eluant solution of methanol/water (V/V) 70:30, collect eluent; Concentrate to get medicinal extract, upper HPLC post separates, 75% methanol aqueous solution wash-out, and the uncommon semi-preparative column of last Japanese aestheticism carries out purifying, and the methanol aqueous solution wash-out with 75% obtains the sweroside compound.
Embodiment 4
Adopt the desugar of D201 macroporous resin column, all the other are with embodiment 1.
Embodiment 5
Adopt the desugar of D301 macroporous resin column, all the other are with embodiment 2.
Embodiment 6
The preparation of the sweroside aqueous solution of 1 μ g/ml.The sweroside compound 2 μ g that take by weighing separating-purifying are dissolved in a small amount of DMSO solution, fully mix, and after it fully dissolves, add the distilled water of surplus to final volume 2ml, make aqua, namely get the sweroside aqueous solution of 1 μ g/ml.Get final product according to the desired concn dilution during use.
Embodiment 7
The preparation of the sweroside aqueous solution of 20 μ g/ml.The sweroside compound 20 μ g that take by weighing separating-purifying are dissolved in a small amount of DMSO solution, fully mix, and after it fully dissolves, add the distilled water of surplus to final volume 1ml, make aqua, namely get the sweroside aqueous solution of 20 μ g/ml.Get final product according to the desired concn dilution during use.
Embodiment 8
The preparation of the 100 μ g/ml sweroside aqueous solution.The sweroside compound 100 μ g that take by weighing separating-purifying are dissolved in a small amount of DMSO solution, fully mix, and after it fully dissolves, add the distilled water of surplus to final volume 1ml, make aqua, namely get the sweroside aqueous solution of 100 μ g/ml.Get final product according to the desired concn dilution during use.
Embodiment 9
Preparation 1g sweroside pulvis takes by weighing the 10mg sweroside, adds a little DMSO and makes it abundant dissolving; And then add 0.1g bovine serum albumin white powder, and add the abundant stirring and evenly mixing of water, then add 20% starch as excipient, fully mixing grinds after the drying and makes pulvis.During use Dilution for powder to desired concn is got final product.
Embodiment 10
Preparation 1g sweroside pulvis takes by weighing the 0.95g sweroside, adds a little DMSO and makes it abundant dissolving; And then add 0.01g bovine serum albumin white powder, and add the abundant stirring and evenly mixing of water, then add surplus starch as excipient, fully mixing grinds after the drying and makes pulvis.During use Dilution for powder to desired concn is got final product.
Embodiment 11
Preparation 1g sweroside granule takes by weighing the 0.15g sweroside, adds a little DMSO and makes it abundant dissolving; And then add 0.5g bovine serum albumin white powder, and add the abundant stirring and evenly mixing of water, then add 15% starch and 15% pectin as excipient, fully mixing becomes granule after granulation and the drying.During use granule being diluted to desired concn gets final product.
Embodiment 12
Preparation 1g sweroside granule takes by weighing the 0.001g sweroside, adds a little DMSO and makes it abundant dissolving; And then add 0.5g bovine serum albumin white powder, and add the abundant stirring and evenly mixing of water, then add 20% starch and 20% pectin as excipient, fully mixing becomes granule after granulation and the drying.During use granule being diluted to desired concn gets final product.
Embodiment 13
Preparation 1g sweroside emulsion takes by weighing the 0.99g sweroside, adds a little DMSO and makes it abundant dissolving; Then add 10% neopelex as emulsifier, fully mixing becomes emulsion.During use emulsion being diluted to desired concn gets final product.
Embodiment 14
Preparation 1g sweroside tablet takes by weighing the 0.4g sweroside, adds a little DMSO and makes it abundant dissolving; And then add 0.1g wheat gluten (powder), and add the abundant stirring and evenly mixing of water, then add 40% starch as excipient, fully mixing is suppressed into tablet after the drying.During use tablet being diluted to desired concn gets final product.
Embodiment 15
The TMV solution preparation of 10 μ g/ml.Draw the TMV mother liquor 1ml of the 100 μ g/ml that prepared, add the phosphate buffer of surplus to final volume 10ml, namely get the TMV solution of 10 μ g/ml.
Embodiment 16
The inhibitory action test that sweroside just infects TMV.Adopt the withered spot method of half leaf to Nicotiana glutinosa artificial infection TMV.Choose healthy and well-grown Nicotiana glutinosa, place a night in the darkroom.Every cigarette seedling is selected 3 identical blades of horizontal level.The right half part of each blade is evenly sprayed the sweroside solution 100 μ l of 10 μ g/ml, and left-half is sprayed the distilled water that 100 μ l contain identical DMSO concentration, the sprinkling amount all soak with the blade face and not lower be as the criterion.Behind 2 h with the TMV 200 μ l of the even frictional inoculation 10 μ g/ml of whole blade.Take 1000 times of liquid of 8% Ningnanmycin aqua as contrast.Each processes 3 strain cigarettes totally 10 tobacco leaves.Add up respectively the withered spot number order of left and right sides blade after 5 days and calculate withered spot inhibiting rate according to the withered spot number on the blade.The inhibiting rate that compound infects TMV=[(positive control withered spot number-compound treatment withered spot number)/positive control withered spot number] * 100%.
Experimental result: table 1 for the compound sweroside when 10 μ g/ml concentration and the rejection ratio that on Nicotiana glutinosa, TMV just infected of control drug Ningnanmycin.By the result as can be known, the compound sweroside is 48.5% to the average inhibiting rate of TMV, and Ningnanmycin only has 35% to the inhibiting rate of TMV.Therefore, sweroside inhibition that TMV is just infected is better than marketed drugs " Ningnanmycin ".
The comparison of the inhibiting rate that table 1 sweroside and Ningnanmycin just infect TMV on Nicotiana glutinosa
Embodiment 17
The sweroside solution of variable concentrations is on the value-added impact test of TMV.Choose healthy and well-grown system host's tobacco bred K326, place a night in the darkroom.Sweroside solution with variable concentrations evenly is sprayed on the blade face, the sprinkling amount all soak with the blade face and not lower be as the criterion.Behind the 2h with the TMV 200 μ l of the even frictional inoculation 10 μ g/ml of whole blade.Take 1000 times of liquid of 8% Ningnanmycin aqua as contrast.Inoculate the propagation situation that detects TMV virus after 3 days with TAS-ELISA.
Experimental result: along with the increase of sweroside concentration, the content of virion reduces gradually.When sweroside concentration was 10 μ g/ml, it obviously was better than the marketed drugs Ningnanmycin to the inhibition that TMV copies in the host.When the concentration of sweroside reaches 15 μ g/ml, can't detect exist (Fig. 1) of virus.
Embodiment 18
Sweroside forms the impact test of symptom at the cigarette seedling on TMV.Choose healthy and well-grown withered spot host Nicotiana glutinosa, place a night in the darkroom.Evenly be sprayed on right half blade face, blade face with the sweroside solution of 15 μ g/ml, left half foliage spray clear water compares, the sprinkling amount all soak with the blade face and not lower be as the criterion.Behind the 2h with the TMV 200 μ l of the even frictional inoculation 10 μ g/ml of whole blade.Take 1000 times of liquid of 8% Ningnanmycin aqua as contrast.The symptom of observing the cigarette seedling on the 5th day behind the inoculation TMV.
Experimental result: as shown in Figure 2, with inoculating TMV behind the 15 μ g/ml sweroside solution-treated cigarette seedlings, substantially do not have withered spot symptom to occur after 5 days on the cigarette seedling.The withered spot number order significantly is less than positive control, also obviously is less than the cigarette seedling that Ningnanmycin is processed.Show that sweroside can suppress or delay the performance of TMV symptom on tobacco, its inhibition is better than the marketed drugs Ningnanmycin.
Claims (10)
1. the application of sweroside pharmaceutical preparation in the control tobacco mosaic virus disease.
2. application as claimed in claim 1 is characterized in that: described sweroside pharmaceutical preparation contains sweroside 0.1 ~ 99%, and all the other are acceptable pharmaceutic adjuvant and/or excipient on the pharmacology.
3. application as claimed in claim 1 or 2 is characterized in that: described sweroside pharmaceutical preparation contains sweroside 0.5 ~ 90%, and all the other are acceptable pharmaceutic adjuvant and/or excipient on the pharmacology.
4. application as claimed in claim 1 or 2 is characterized in that: described sweroside pharmaceutical preparation contains sweroside 1 ~ 50%, and all the other are acceptable pharmaceutic adjuvant and/or excipient on the pharmacology.
5. application as claimed in claim 1 or 2 is characterized in that: described sweroside pharmaceutical preparation contains sweroside 5 ~ 20%, and all the other are acceptable pharmaceutic adjuvant and/or excipient on the pharmacology.
6. application as claimed in claim 1 or 2 is characterized in that: the sweroside in the described sweroside pharmaceutical preparation is take surrounded swertia herb as raw material, and extracts by the technique that comprises the following steps:
A, get the surrounded swertia herb that dries in the shade and pulverize, with 75 ~ 85% methyl alcohol extraction 2 ~ 3 times, each 1 ~ 3h;
B, merging extract, filter cleaner, merging filtrate, then reduced pressure concentration gets the medicinal extract of relative density 1.2 ~ 1.3;
The water of C, 1 ~ 3 times of volume of medicinal extract adding is stirred to and is suspension, with concentration 95 ~ 100% dichloromethane solutions extraction 2 ~ 3 times, each 15 ~ 25min;
D, combining extraction liquid are crossed the decolouring of MCI post, use afterwards the methanol-eluted fractions of concentration 70 ~ 80%, collect eluent;
E, use the macroporous resin column desugar again, and with the eluant solution of methanol/water (V/V) 0 ~ 100:100 ~ 0, collect eluent, concentrated the medicinal extract that contains 70 ~ 80% methyl alcohol;
F, upper HPLC separate, and with 70 ~ 80% methanol aqueous solution wash-outs, carry out purifying with semi-preparative column at last, obtain the sweroside compound.
7. application as claimed in claim 6 is characterized in that: described macroporous resin column is any one among D900, D130, D101, D201 or the D301.
8. application as claimed in claim 6 is characterized in that: described semi-preparative column is any one in the uncommon semi-preparative column of Ai Jieer semi-preparative column, HPLC semi-preparative column or Japanese aestheticism.
9. application as claimed in claim 1 or 2, it is characterized in that: described sweroside pharmaceutical preparation with the sweroside compound and to the nontoxic pharmaceutically acceptable auxiliaries of humans and animals and/or mixed with excipients, is mixed with aqua, tablet, granule or microemulsion by prescription requirement ratio.
10. application as claimed in claim 1 or 2 is characterized in that: the sweroside concentration of described sweroside pharmaceutical preparation when preventing and treating tobacco mosaic is 1 ~ 20 μ g/ml.
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| CN104910224A (en) * | 2015-07-04 | 2015-09-16 | 云南省农业科学院生物技术与种质资源研究所 | Iridoid compound, preparation method, preparation and appliance thereof |
| CN105017354B (en) * | 2015-07-04 | 2018-06-19 | 云南民族大学 | A kind of iridoid and preparation method thereof, preparation and application |
| CN107573392B (en) * | 2017-10-16 | 2020-12-22 | 南开大学 | Glycosyl-substituted genipin derivative and preparation and application thereof |
| CN113577131A (en) * | 2021-06-29 | 2021-11-02 | 内蒙古民族大学 | Application of Mongolian medicine extract of lomatogonium carinthiacum |
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