CN106038725A - Method for separating anti-RSV-virus effective constituents after compatibility of pulsatilla chinensis and holotrichia diomphalia extracts - Google Patents
Method for separating anti-RSV-virus effective constituents after compatibility of pulsatilla chinensis and holotrichia diomphalia extracts Download PDFInfo
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- CN106038725A CN106038725A CN201610558451.6A CN201610558451A CN106038725A CN 106038725 A CN106038725 A CN 106038725A CN 201610558451 A CN201610558451 A CN 201610558451A CN 106038725 A CN106038725 A CN 106038725A
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- 241001465967 Holotrichia diomphalia Species 0.000 title claims abstract description 23
- 239000000284 extract Substances 0.000 title claims abstract description 10
- 238000000034 method Methods 0.000 title claims abstract description 8
- 241000123887 Pulsatilla chinensis Species 0.000 title abstract 5
- 239000000470 constituent Substances 0.000 title abstract 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 26
- 230000000840 anti-viral effect Effects 0.000 claims abstract description 20
- 238000000605 extraction Methods 0.000 claims abstract description 19
- 239000011347 resin Substances 0.000 claims abstract description 13
- 229920005989 resin Polymers 0.000 claims abstract description 13
- 238000000926 separation method Methods 0.000 claims abstract description 13
- 238000001556 precipitation Methods 0.000 claims abstract description 12
- 230000000694 effects Effects 0.000 claims abstract description 9
- 230000002155 anti-virotic effect Effects 0.000 claims abstract description 8
- 241000700605 Viruses Species 0.000 claims abstract description 7
- 238000010992 reflux Methods 0.000 claims abstract description 4
- 238000011160 research Methods 0.000 claims abstract description 3
- 239000009806 pulsatillae Substances 0.000 claims description 22
- 238000009395 breeding Methods 0.000 claims description 5
- 230000001488 breeding effect Effects 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 238000002474 experimental method Methods 0.000 claims description 4
- 239000004615 ingredient Substances 0.000 claims description 4
- 239000003480 eluent Substances 0.000 claims description 3
- 238000004080 punching Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 201000010099 disease Diseases 0.000 claims description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 231100000614 poison Toxicity 0.000 claims description 2
- 238000000746 purification Methods 0.000 claims description 2
- 239000002244 precipitate Substances 0.000 claims 3
- 239000006286 aqueous extract Substances 0.000 claims 1
- -1 is 1:10 Substances 0.000 claims 1
- 239000002574 poison Substances 0.000 claims 1
- 230000001629 suppression Effects 0.000 claims 1
- 239000012153 distilled water Substances 0.000 abstract description 5
- 239000003814 drug Substances 0.000 abstract description 5
- 238000004090 dissolution Methods 0.000 abstract description 3
- 239000004480 active ingredient Substances 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 abstract description 2
- 238000001179 sorption measurement Methods 0.000 abstract 2
- 239000000243 solution Substances 0.000 description 5
- 241000725643 Respiratory syncytial virus Species 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000003463 adsorbent Substances 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 239000012567 medical material Substances 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 206010061603 Respiratory syncytial virus infection Diseases 0.000 description 2
- IWUCXVSUMQZMFG-AFCXAGJDSA-N Ribavirin Chemical compound N1=C(C(=O)N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 IWUCXVSUMQZMFG-AFCXAGJDSA-N 0.000 description 2
- 239000003443 antiviral agent Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 229960000329 ribavirin Drugs 0.000 description 2
- HZCAHMRRMINHDJ-DBRKOABJSA-N ribavirin Natural products O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1N=CN=C1 HZCAHMRRMINHDJ-DBRKOABJSA-N 0.000 description 2
- 238000007873 sieving Methods 0.000 description 2
- 208000008710 Amebic Dysentery Diseases 0.000 description 1
- 206010001986 Amoebic dysentery Diseases 0.000 description 1
- 206010006448 Bronchiolitis Diseases 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 206010061598 Immunodeficiency Diseases 0.000 description 1
- 208000029462 Immunodeficiency disease Diseases 0.000 description 1
- 208000029523 Interstitial Lung disease Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000475481 Nebula Species 0.000 description 1
- 208000022873 Ocular disease Diseases 0.000 description 1
- 206010068319 Oropharyngeal pain Diseases 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 208000003251 Pruritus Diseases 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- 241000315672 SARS coronavirus Species 0.000 description 1
- 241000144290 Sigmodon hispidus Species 0.000 description 1
- 206010043376 Tetanus Diseases 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000003097 anti-respiratory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 208000002399 aphthous stomatitis Diseases 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 208000001848 dysentery Diseases 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 231100000734 genotoxic potential Toxicity 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 230000007813 immunodeficiency Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 238000011552 rat model Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 208000030925 respiratory syncytial virus infectious disease Diseases 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/63—Arthropods
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
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- Animal Behavior & Ethology (AREA)
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- Engineering & Computer Science (AREA)
- Insects & Arthropods (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
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- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to traditional Chinese medicine active ingredient extraction process and antiviral effective part separation, and mainly relates to a method for separating anti-RSV-virus effective constituents after compatibility of pulsatilla chinensis and holotrichia diomphalia extracts. The method particularly comprises the following steps: conducting reflux extraction on pulsatilla chinensis and holotrichia diomphalia by adopting distilled water, conducting alcohol precipitation, resisting viruses, conducting dissolution and precipitation, separating the pulsatilla chinensis extract by adopting X-5 type macroporous adsorption resin to achieve a better effect, separating the holotrichia diomphalia extract by adopting D101 type macroporous adsorption resin to achieve a better effect, and determining the anti-virus effective parts. Then the effective parts of the two medicines are in compatibility according to the proportion of 9:1, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8 and 1:9 respectively, so as to conduct anti-RSV-virus research again. Through adoption of the anti-virus tracing method, the fact that after the compatibility of the anti-virus effective parts of the pulsatilla chinensis and the holotrichia diomphalia, the anti-virus effect is better, and the optimal compatibility proportion is obtained.
Description
Technical field
The present invention relates to active ingredient of Chinese herbs and produce extraction process, relate to the Radix Pulsatillae, the extraction of Holotrichia diomphalia Bates antivirus effective position
New approaches new method and at the new application of pharmaceutical field, particularly relate to the mode extraction separation and purification that antiviral is followed the trail of
The Radix Pulsatillae, the effective ingredient of Holotrichia diomphalia Bates anti respiratory syncytial virus, and effective ingredient carry out the antiviral study after compatibility.
Background technology
In recent years due to the unexpected invasion of various viruses, such as SARS virus, H1N1 etc., each time outburst of virus
The fear that will cause the mankind is the most dead, so finding suitable antiviral drugs most important.
The Radix Pulsatillae is the dry root of the Mao Moke plant Radix Pulsatillae, has another name called old man and spends, Bai Toucao, Radix Glycyrrhizae, is always preced with flower, is born in mountain
Between open country, barren hill and field, it is distributed in the ground such as northeast, North China, northwest, East China.
The Radix Pulsatillae begins to be loaded in " herbal classic ", has effect of heat-clearing and toxic substances removing, eliminating pathogenic heat from blood to cure dysentery, can be used for treating toxic-heat and blood stasis, the moon
Itch leukorrhagia, amebic dysentery etc..
Respiratory syncytial virus is to cause one of most important pathogen of infant acute lower respiratory infection, also can cause
Old people and immunodeficiency patient can also be caused to be in hospital simultaneously for interstitial pneumonia and bronchiolitis etc. and pneumonia is dead.
Ribavirin is the antiviral drugs having certain curative effect to respiratory syncytial virus of wide spectrum.But, research finds,
Cotton rat model through respiratory syncytial virus infection occurs in that the resistant strain of ribavirin, and has patient and medical worker
Genotoxic potential, so only occurring serious rsv infection patient is just to use this medicine, therefore explores respiratory syncytial virus
Inhibited novel substance is required by the epoch.
Holotrichia diomphalia Bates has long use history in motherland's traditional medicine.There are the effects such as removing blood stasis, clots absorbing, eliminating stagnation, lactogenesis, use
The diseases such as stasis of blood pain, tetanus, sore throat, corneal nebula are lost in treatment.
In terms of pharmacology, grub extract has antitumor, the treatment multiple biological activity such as aphtha and ocular disease, is
The animal drugs that a kind of great research and development is worth.
Summary of the invention
The technical problem to be solved is to specify that the Radix Pulsatillae and the effective site of Holotrichia diomphalia Bates anti-RSV virus, for this
The separation and Extraction of effective site provides a kind of mode utilizing antiviral to follow the trail of and prepares the Radix Pulsatillae, Holotrichia diomphalia Bates antivirus effective position
New method, and find both optimal proportion compatibilities.
It is an advantage of the invention that the present invention to isolate first in the Radix Pulsatillae, Holotrichia diomphalia Bates and there is a kind of effective of anti-RSV virus
Position, and invented a kind of method that antiviral follows the trail of prepared by this effective site, this separation method purposiveness is strong, letter
Singly save substantial amounts of manpower and materials, be better than traditional system and separate.RSV virus is had preferably by the effective site prepared
Therapeutical effect.
The Radix Pulsatillae of the present invention and the optimal proportion compatibility of Holotrichia diomphalia Bates effective site antiviral are 2:3 position and optimal proportion compatibility
Preparation method, comprise the steps:
(1) a certain amount of Radix Pulsatillae, Holotrichia diomphalia Bates medical material are taken respectively by the water heating and refluxing extraction 2 times of 10 times, 2 hours second for the first time
Secondary 1.5 hours.
(2) merge the extracting solution concentrating under reduced pressure of twice, be concentrated into the medical material that every milliliter of medicinal liquid contains 1 gram, then add ethanol
The concentration of regulation ethanol is 70%, stands 24 hours.It is filtrated to get precipitation and supernatant.
(3) supernatant is reclaimed ethanol, by precipitation water dissolution, carry out Antiviral breeding respectively.Experiment display precipitation
Antiviral effect is preferable.
(4) take certain D101, X-5, HPD100, DM130, AB-8 model macroporous adsorbent resin respectively with 95% second
Alcohol soaks 24 hours, fills post afterwards, is washed till without alcohol taste with distilled water, then rushes with the aqueous hydrochloric acid solution of 5 times of column volumes 5%
Wash immersion 3 hours, afterwards with distilled water punching to PH equal to 7, then be rinsed leaching with the NaOH aqueous solution of the 5% of 5 times of column volumes
Steep 3 hours, afterwards with distilled water punching to PH equal to 7, standby.
(5) take the precipitation in (3) and be splined on the macroporous resin column handled well, respectively with water, 25% ethanol, 50% ethanol, 75%
Ethanol is rinsed, and in each eluent, every 200ml connects one bottle, and changes next eluting after being flushed to not have color again
Liquid.
(6) the above-mentioned sample received is carried out Antiviral breeding.Experiment display is through the screening of macroporous resin, the Radix Pulsatillae pair
The effect of X-5 macroporous resin is preferable, and Holotrichia diomphalia Bates is to the D101 preferable Radix Pulsatillae of macroporous adsorbent resin effect and Holotrichia diomphalia Bates water position antiviral
Effect is fine.
(7) by the water position of the Radix Pulsatillae and Holotrichia diomphalia Bates respectively with 9:1,8:2,7:3,6:4,5:5,4:6,3:7,2:8,1:9
Ratio carries out compatibility, and result displaying ratio 2:3 is optimal proportion compatibility.
Detailed description of the invention
Example 1(1) weigh a certain amount of Radix Pulsatillae, Holotrichia diomphalia Bates and add 10 times amount heating and refluxing extraction respectively twice, 2 hours for the first time,
1.5 hours for the second time, merge twice extracting solution, be evaporated to the medical material that every milliliter of medicinal liquid contains 1 gram, add ethanol regulation solution
The concentration of ethanol is 70%, stands 24 hours.
(2) the alcohol deposit fluid decompression sucking filtration (1) obtained, is precipitated, precipitation is used distilled water ultrasonic dissolution.
(3) solution that (2) obtain is splined on X-5, D101 macroporous adsorbent resin handled well, is rinsed with water, often
200ml connects one bottle, rinses to colourless.
(4) merge water elution position in (3), be concentrated into certain volume, carry out antiviral study respectively.
(5) by the Radix Pulsatillae water elution position through X-5 Resin sieving selection obtained in (4) and through D101 Resin sieving selection
Holotrichia diomphalia Bates water elution position water eluting position is joined with the ratio of 9:1,8:2,7:3,6:4,5:5,4:6,3:7,2:8,1:9 respectively
5, again carry out Antiviral breeding, after the result display Radix Pulsatillae and Holotrichia diomphalia Bates compatibility, antiviral effect is preferable, and ratio 2:3 is
Optimal proportion compatibility.
Claims (10)
1. the Radix Pulsatillae and grub extract have the effective ingredient of anti-RSV virus function after compatibility and one utilizes antiviral
The process of mode this effective ingredient of extraction separation and purification followed the trail of, it is characterised in that the Radix Pulsatillae, the water extract-alcohol precipitation of Holotrichia diomphalia Bates
Precipitation position has the effect of suppression RSV virus, and first with water extract-alcohol precipitation, next utilizes the macroporous resin of X-5, D101 model
Separating, after the extraction of each step separates, antiviral to be carried out tracks out the place of effective site, then will have antivirus action
Position carry out compatibility, finally carry out Antiviral breeding research, draw optimal compatibility program.
Effective site the most according to claim 1 is characterized in that the precipitation position that this effective site is water extract-alcohol precipitation, warp
After macroporous resin separates, water position is the effective site of the Radix Pulsatillae and Holotrichia diomphalia Bates antiviral, and the optimal proportion compatibility at water position is 2:
3。
The most according to claim 1, extraction separation method, it is characterised in that the solid-liquid ratio of water reflux, extract, is 1:10, extract secondary
Several twice, extraction time is 2 hours for the first time, 1.5 hours for the second time.
Extraction separation method the most according to claim 1, it is characterised in that the concentrated solution concentration of Aqueous extracts is 1g/ml, precipitate with ethanol
Concentration is 70%, and the precipitate with ethanol time is 24 hours.
Extraction separation method the most according to claim 1, it is characterised in that carry out after precipitate with ethanol reduce pressure sucking filtration, be precipitated and
Supernatant, then carries out Antiviral breeding and is precipitated as antivirus effective position.
Extraction separation method the most according to claim 1, it is characterised in that macroporous resin model used is X-5, D101, washes
De-liquid is water, 25% ethanol, 50% ethanol, and the 75% every 200ml of ethanol connects one bottle, and the punching of each gradient is to colourless.
Extraction separation method the most according to claim 1, it is characterised in that the eluent of Radix Pulsatillae extract is carried out disease-resistant
Poison experiment, obtaining water position is effective site.
Extraction separation method the most according to claim 1, it is characterised in that the eluent of grub extract is carried out antiviral
Experiment, obtaining water position is effective site.
Extraction separation method the most according to claim 1, it is characterised in that by Radix Pulsatillae water position with Holotrichia diomphalia Bates water position by one
Certainty ratio compatibility, proportion compatibility is respectively 9:1,8:2,7:3,6:4,5:5,4:6,3:7,2:8,1:9.
Extraction separation method the most according to claim 1, it is characterised in that optimal antiviral proportion compatibility is 2:3.
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| CN201610558451.6A CN106038725A (en) | 2016-07-16 | 2016-07-16 | Method for separating anti-RSV-virus effective constituents after compatibility of pulsatilla chinensis and holotrichia diomphalia extracts |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610558451.6A CN106038725A (en) | 2016-07-16 | 2016-07-16 | Method for separating anti-RSV-virus effective constituents after compatibility of pulsatilla chinensis and holotrichia diomphalia extracts |
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| CN106038725A true CN106038725A (en) | 2016-10-26 |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1471914A (en) * | 2003-04-29 | 2004-02-04 | 胡世玺 | Medicine, application and preparing method for preventnig SARS pneumonia by SARS virus |
| CN1824080A (en) * | 2005-12-27 | 2006-08-30 | 广东大光药业有限公司 | Medicinal composition for antivirus and its preparation method |
| CN101023976A (en) * | 2006-10-26 | 2007-08-29 | 正大青春宝药业有限公司 | Medicine composition with anti-virus function and its preparing method |
| CN101780239A (en) * | 2009-01-15 | 2010-07-21 | 颜平安 | Four-herb anti-tumor and anti-AIDS soft capsules and preparation process thereof |
| CN104803851A (en) * | 2015-02-26 | 2015-07-29 | 周长征 | Mint effective ingredient capable of resisting respiratory syncytial virus and preparation method |
-
2016
- 2016-07-16 CN CN201610558451.6A patent/CN106038725A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1471914A (en) * | 2003-04-29 | 2004-02-04 | 胡世玺 | Medicine, application and preparing method for preventnig SARS pneumonia by SARS virus |
| CN1824080A (en) * | 2005-12-27 | 2006-08-30 | 广东大光药业有限公司 | Medicinal composition for antivirus and its preparation method |
| CN101023976A (en) * | 2006-10-26 | 2007-08-29 | 正大青春宝药业有限公司 | Medicine composition with anti-virus function and its preparing method |
| CN101780239A (en) * | 2009-01-15 | 2010-07-21 | 颜平安 | Four-herb anti-tumor and anti-AIDS soft capsules and preparation process thereof |
| CN104803851A (en) * | 2015-02-26 | 2015-07-29 | 周长征 | Mint effective ingredient capable of resisting respiratory syncytial virus and preparation method |
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