CN104803851A - Mint effective ingredient capable of resisting respiratory syncytial virus and preparation method - Google Patents
Mint effective ingredient capable of resisting respiratory syncytial virus and preparation method Download PDFInfo
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- CN104803851A CN104803851A CN201510088480.6A CN201510088480A CN104803851A CN 104803851 A CN104803851 A CN 104803851A CN 201510088480 A CN201510088480 A CN 201510088480A CN 104803851 A CN104803851 A CN 104803851A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
- C07C69/02—Esters of acyclic saturated monocarboxylic acids having the carboxyl group bound to an acyclic carbon atom or to hydrogen
- C07C69/12—Acetic acid esters
- C07C69/14—Acetic acid esters of monohydroxylic compounds
- C07C69/145—Acetic acid esters of monohydroxylic compounds of unsaturated alcohols
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- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the field of production and extraction technologies and structural identification of active ingredients of Chinese medicine, and relates to a novel technology for identification of a mint effective ingredient resisting to a respiratory syncytial virus and extraction and separation of the effective ingredient as well as a novel application of the effective ingredient in the pharmacy field, in particular to extraction and preparation of the mint effective ingredient resisting to the respiratory syncytial virus in an antivirus tracking manner as well as identification of the effective ingredient with a mass spectrum nuclear magnetism technology. The method comprises specific steps of reflux extraction of mint by the aid of distilled water, alcohol precipitation, virus resistance, dissolution precipitation, treatment of D101 macroporous resin, sample loading in the macroporous resin and different-concentration gradient elution, wherein every 200 ml of each concentration gradient eluent is contained in one bottle and washed to be colorless, each eluent is subjected to anti-virus treatment to determine parts, the parts are observed under an ultraviolet lamp, effective parts are merged, a thin layer is prepared, an anti-virus experiment is performed, the effective part is determined, and the ingredient in the effective part is identified with the mass spectrum nuclear magnetism technology. With adoption of an anti-virus tracking method, the mint effective ingredient resisting to the respiratory syncytial virus is firstly found out.
Description
Technical field
The present invention relates to active ingredient of Chinese herbs production extraction process and Structural Identification field, relate to the new approaches novel method of peppermint antivirus effective position extraction and the novelty teabag at pharmacy field, be specifically related to the effective constituent of the mode extraction separation and purification peppermint anti respiratory syncytial virus utilizing antiviral tracking, and this effective constituent carries out Structural Identification and application.
Background technology
All receive the invasion of various virus since the last few years all over the world, such as SARS virus, H1N1 etc., the outburst of each virus all can cause fear and the death of the mankind.So find suitable antiviral extremely urgent.
Respiratory syncytial virus causes one of most important pathogenic agent of infant's acute lower respiratory infection, and interstitial pneumonia and capillary bronchitis etc. also can be caused the elderly and immune deficiency patient also can be caused to be in hospital and pneumonia death simultaneously.
Respiratory syncytial virus just received much concern before 50 years, but still lacked its effective prophylactic treatment measure at present.Ribavirin is antiviral respiratory syncytial virus being had to certain curative effect of wide spectrum.But, research finds, there is the resistant strain of ribavirin in the cotton rat model through respiratory syncytial virus infection, and there is genotoxic potential to patient and medical worker, so only occur serious rsv infection patient just use this medicine, therefore explore the inhibited novel substance of respiratory syncytial virus as the epoch are required.
Peppermint is the dry aerial parts of labiate peppermint Mentha haplocalyx Briq..Be usually used in common cold due to wind-heat, wind-warm syndrome from the beginning of, headache, hot eyes, larynx numbness, aphtha, rubella, measles, chest the side of body swollen vexed.Peppermint is Chinese medicine compound prescription Lonicerae and Forsythiae Powder, the composition medicine of GANMAO QINGRE KELI.Lonicerae and Forsythiae Powder is the clinical the most frequently used prescription of warm disease, is widely used in the initial stage of the various diseases for the treatment of within the scope of warm disease.GANMAO QINGRE KELI be the common drug for the treatment of common cold caused by wind-cold be usually used in clinically treatment cold, fever, cough headache, swelling and pain in the throat and measles from the beginning of etc. disease.
Peppermint has certain therapeutic action to respiratory syncytial virus, and experiment shows that the effective constituent that peppermint is extracted has good therapeutic action to respiratory syncytial virus.
Summary of the invention
Technical problem to be solved by this invention is the effective constituent that specify that peppermint anti respiratory syncytial virus, and provides a kind of novel method utilizing the mode of antiviral tracking to prepare the antiviral effective constituent of peppermint for the separation and Extraction of this effective constituent.This effective constituent extraction and isolation from labiate peppermint obtains the treatment that may be used for respiratory syncytial virus infection.The method can be comparatively easy separation and Extraction is carried out to above effective constituent.
Advantage of the present invention is, the present invention has isolated in peppermint a kind of effective constituent with anti respiratory syncytial virus first, and a kind of method of having invented antiviral tracking is prepared this effective constituent, this separation method purpose is strong, simply save a large amount of manpower and materials, be better than traditional systematic position.The effective constituent prepared has good therapeutic action to respiratory syncytial virus.
The effective constituent of peppermint anti respiratory syncytial virus of the present invention is 3-acetoxyl group-p-menthane-4,8-alkene or 3-acetoxyl group-p-menthane-1,2-alkene, and the preparation method of this effective constituent, comprises the steps:
(1) get a certain amount of peppermint medicinal material water heating and refluxing extraction 2 times of 10 times, 1.5 hours first times second time 1 hour.
(2) merge the extracting solution concentrating under reduced pressure of twice, be concentrated into the medicinal material that every milliliter of liquid contains 1 gram, then add ethanol and regulate the concentration of ethanol to be 50%, leave standstill 24 hours.Filtration is precipitated and supernatant liquor.
(3) supernatant liquor is reclaimed ethanol, by precipitation water dissolution, carry out Antiviral breeding respectively.The antiviral effect of experiment display precipitation is better.
(4) get certain D101 macroporous adsorbent resin with 95% alcohol immersion 24 hours, fill post afterwards, be washed till without alcohol taste with distilled water, then flushing immersion 3 hours is carried out with the aqueous hydrochloric acid of 5 times of column volumes 5%, 7 are equaled to PH afterwards with distilled water punching, carry out flushing immersion 3 hours with the NaOH aqueous solution of 5% of 5 times of column volumes again, equal 7 with distilled water punching to PH afterwards, for subsequent use.
(5) precipitation of getting in (3) is splined on the D101 macroporous resin column handled well, uses water respectively, 25% ethanol, 50% ethanol, 75% ethanol, 95% ethanol rinses, in each elutriant, every 200ml connects one bottle, and be flushed to there is no color after change next elutriant again.
(6) the above-mentioned sample received is carried out Antiviral breeding.Experiment display water No. 6 No. 7 No. 8 antiviral effects are fine.
(7) by above-mentioned 3 sample spot on the thin layer plate of silica gel G, with developping agent (chloroform: methyl alcohol: water=30: 12: 4, takes off a layer 9ml, adds the Glacial acetic acid of 1.2ml).Be placed under ultraviolet lamp (254nm) and observe, its speckle displacement equally therefore merge 3 elution samples.
(8) method of elution samples Preparative TLC be separated, wherein stationary phase is silica gel G, and developping agent is (chloroform: methyl alcohol: water=30: 12: 4, takes off a layer 9ml, adds the Glacial acetic acid of 1.2ml).
(9) observe under the plank obtained being placed in ultraviolet lamp, collect spot and non-spot, use methyl alcohol ultrasonic dissolution respectively, afterwards evaporate to dryness, use water dissolution.
(10) Antiviral breeding is carried out respectively in the spot (9) obtained and non-spot position, and experimental result display dot and non-spot all have good antiviral effect.
(11) utilized by the spot that (10) obtain mass spectrum and nuclear-magnetism equipment to carry out qualification of setting the chessman on the chessboard according to the chess manual, qualification result for this effective constituent be 3-acetoxyl group-p-menthane-4,8-alkene or 3-acetoxyl group-p-menthane-1,2-alkene
Embodiment
Example 1 (1) takes a certain amount of peppermint and adds 10 times amount heating and refluxing extraction twice, 1.5 hours first times, second time 1 hour, merge extracted twice liquid, be evaporated to the medicinal material that every milliliter of liquid contains 1 gram, the concentration adding ethanol regulator solution ethanol is 50%, leaves standstill 24 hours.
(2) by the precipitation solution decompress filter that (1) obtains, be precipitated, precipitation used distilled water ultrasonic dissolution.
(3) solution that (2) obtain is splined on the D101 macroporous adsorbent resin handled well, rinses with water, every 200ml connects one bottle, rinses to colourless.
(4) 6,7 are merged in (3), No. 8 bottles, be concentrated into certain volume, point sample is in preparing silica gel g thin-layer plate, and wherein developping agent is chloroform: methyl alcohol: water=30: 12: 4, take off a layer 9ml, add the Glacial acetic acid of 1.2ml, under ultraviolet lamp (254nm), observe spot situation.
(5) the spot methyl alcohol ultrasonic dissolution will obtained in (4), is placed in evaporate to dryness on water-bath, dissolves with distilled water.
(6) solution obtained in (5) is carried out antiviral study in vitro and experiment in vivo, it is better that experimental result shows its antiviral effect.
Claims (15)
1. there is in a peppermint effective constituent of anti respiratory syncytial virus effect and a kind of processing method utilizing this effective constituent of mode extraction separation and purification of antiviral tracking, it is characterized in that this effective constituent is 3-acetoxyl group-p-menthane-4, 8-alkene or 3-acetoxyl group-p-menthane-1, 2-alkene, the antiviral place tracking out efficient part to be carried out after the extraction and isolation of each step, first water extract-alcohol precipitation is utilized, next utilizes D101 macroporous resin to be separated, finally separate effective spot with Preparative TLC, then the antiviral efficient part of peppermint is obtained through methanol extraction evaporate to dryness.
2. effective constituent according to claim 1 is characterized in that this effective constituent is 3-acetoxyl group-p-menthane-4,8-alkene or 3-acetoxyl group-p-menthane-1,2-alkene.
3. effective constituent according to claim 1 is characterized in that the data of its nuclear-magnetism are
13c-NMR (DMSO) δ: 177.50 (C-1), 130.92 (C-2), 128.85 (C-3), 65.60 (C-4), 38.74 (C-5), 34.01 (C-6), 30.57 (C-7), 29.70 (C-8), 24.47 (C-9), 23.75 (C-10), 22.96 (C-11).
1H-NMR(DMSO)δ：1.65(1H，S，1-H)，1.40(2H，t，J＝6.0Hz，2-H)，4.22(1H，S，3-H)，1.96(2H，t，J＝6.0Hz，5-H)，1.40(2H，t，J＝6.0Hz，6-H)0.92(3H，s，7-H)，1.64(3H，s，9-H，10-H)，2.27(3H，s，11-H)。
4. extraction and separation method according to claim 1, is characterized in that the solid-liquid ratio of peppermint water refluxing extraction is 1: 10, extraction time twice, and extraction time is 1.5 hours first times, second time 1 hour.
5. extraction and separation method according to claim 1, it is characterized in that the concentrated solution concentration of Aqueous extracts is the medicinal material that 1ml liquid contains 1g, alcohol precipitation concentration is 50%, and the alcohol precipitation time is 24 hours.
6. extraction and separation method according to claim 1, carries out decompress filter after it is characterized in that alcohol precipitation, is precipitated and supernatant liquor, then carry out Antiviral breeding and be precipitated as antivirus effective position.
7. extraction and separation method according to claim 1, it is characterized in that macroporous resin model used is D101, elutriant is water, 25% ethanol, 50% ethanol, 75% ethanol, 95% ethanol, and every 200ml connects one bottle, and each gradient punching is to colourless.
8. extraction and separation method according to claim 1, it is characterized in that elutriant to carry out Antiviral breeding, obtain 6,7, No. 8 is efficient part.
9. extraction and separation method according to claim 1, it is characterized in that 6,7, No. 8 points launch on silica gel g thin-layer plate, observe its spot consistent, therefore can merge under ultraviolet lamp.
10. extraction and separation method according to claim 1, it is characterized in that the condition of Preparative TLC is developping agent chloroform: methyl alcohol: water=30: 12: 4, take off a layer 9ml, add the Glacial acetic acid of 1.2ml, ultraviolet lamp condition is 254nm.
11. extraction and separation method according to claim 1, is characterized in that it is its antiviral efficient part that spot and non-spot are carried out Antiviral breeding determination spot.
12. extraction and separation method according to claim 1, is characterized in that, by spot methyl alcohol ultrasonic dissolution, being placed in evaporate to dryness on water-bath, obtain the antiviral effective constituent of peppermint.
13. effective constituents according to claim 1-3 comprise its derivative if the glycosides that is parent with this active substance etc. is in the application of medical association area.
14. extract according to the arbitrary described technique of claim 4-12 the phytochemicals production obtained.
15. according to the purposes of the arbitrary described technique of claim 4-12 in effective ingredients in plant extraction field.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201510088480.6A CN104803851A (en) | 2015-02-26 | 2015-02-26 | Mint effective ingredient capable of resisting respiratory syncytial virus and preparation method |
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| CN201510088480.6A CN104803851A (en) | 2015-02-26 | 2015-02-26 | Mint effective ingredient capable of resisting respiratory syncytial virus and preparation method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105998082A (en) * | 2016-07-16 | 2016-10-12 | 周长征 | Effective part capable of resisting respiratory syncytial viruses in grubs and preparation method |
| CN106038725A (en) * | 2016-07-16 | 2016-10-26 | 周长征 | Method for separating anti-RSV-virus effective constituents after compatibility of pulsatilla chinensis and holotrichia diomphalia extracts |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102319287A (en) * | 2011-08-11 | 2012-01-18 | 刘斌 | Mentha haplocalyx phenolic acid extraction and application in preparation of medicines for resisting respiratory syncytial virus |
-
2015
- 2015-02-26 CN CN201510088480.6A patent/CN104803851A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102319287A (en) * | 2011-08-11 | 2012-01-18 | 刘斌 | Mentha haplocalyx phenolic acid extraction and application in preparation of medicines for resisting respiratory syncytial virus |
Non-Patent Citations (3)
| Title |
|---|
| JEAN-CLAIDE FIAUD ET AL: "Palladium-catalyzed synthesis of substituted cyclopentadienes and indenes from allylic estres", 《JOURNAL OF ORGANOMETALLIC CHEMISTRY》 * |
| V. K. AGNIHOTRI ET AL: "Essential oil composition of Mentha pulegium L. growing wild in the north-western Himalayas India", 《FLAVOUR AND FRAGRANCE JOURNAL》 * |
| 姚梅悦等: "感冒清热颗粒体外抗病毒有效成分研究", 《药物研究》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105998082A (en) * | 2016-07-16 | 2016-10-12 | 周长征 | Effective part capable of resisting respiratory syncytial viruses in grubs and preparation method |
| CN106038725A (en) * | 2016-07-16 | 2016-10-26 | 周长征 | Method for separating anti-RSV-virus effective constituents after compatibility of pulsatilla chinensis and holotrichia diomphalia extracts |
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Application publication date: 20150729 |