CN101347492A - Use of Trigonostemon xyphophylloides extract in preparing anti-AIDS medicament - Google Patents
Use of Trigonostemon xyphophylloides extract in preparing anti-AIDS medicament Download PDFInfo
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- CN101347492A CN101347492A CNA2008102135031A CN200810213503A CN101347492A CN 101347492 A CN101347492 A CN 101347492A CN A2008102135031 A CNA2008102135031 A CN A2008102135031A CN 200810213503 A CN200810213503 A CN 200810213503A CN 101347492 A CN101347492 A CN 101347492A
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- xyphophylloides
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Abstract
The present invention relates to a biological medicine, in particular to the application of an extract of xiphophyllous trigonostemon in preparing anti-ADIS drugs. The extract is used for preparing anti-ADIS drugs. The extract of xiphophyllous trigonostemon is the plant extract of xiphophyllous trigonostemon through organic solvent extraction. The extraction process includes baking and grinding xiphophyllous trigonostemon into powder, drying in vacuum until the weight is constant, refluxing and extracting by organic solvent for three times, combining the extract, filtering, reducing pressure and recovering all the solvent, and frozen drying to form powder, thus obtaining the extract of xiphophyllous trigonostemon. The application of the present invention has the advantages of high biological utilization rate of plants, obvious effect, special material source and high safety and has profound significance to deep research and development of the medicine activity of the extract and wide development prospect.
Description
Technical field
The present invention relates to biological medicine, be specifically related to the application of a kind of Trigonostemon xyphophylloides extract in preparation AIDS virus resisting medicine.
Background technology
Infected and the disease that causes is called acquired immune deficiency syndrome (AIDS) by HIV, full name is that (acquired immunodeficiency syndrome, AIDS), this patient's immunologic function partly or completely lose CD4 for acquired immune deficiency syndrome (AIDS)
+Cell number reduces, generator opportunistic infections, tumor etc. then, and clinical manifestation is varied.This disease spread speed is fast, case fatality rate is high, and can't cure at present, has caused national governments and social concerns.
Find two kinds of HIV at present, be respectively HIV-1 and HIV-2.Both have similar virus structure and route of transmission.It is western that HIV-2 mainly is distributed in Africa, also is detected in some the infecteds in Europe and America.Its virulence and transmissibility all are lower than HIV-1, the slow and mitigation of the acquired immune deficiency syndrome (AIDS) course of disease that causes.HIV-1 is distributed widely in all over the world, is to cause the popular cause of disease of whole world AIDS, and the research of HIV at present also is main carrying out with HIV-1.At present the effective method of treatment acquired immune deficiency syndrome (AIDS) is highly active antiretroviral therapy (Highly activeanti-retroviral therapy, HAART, be commonly called as HAART), it is by using 3-4 kind medicine simultaneously, each medicine is at the different links of HIV (human immunodeficiency virus) (Human Immunodeficiency Virus) in the breeding cycle, thereby reach inhibition or kill HIV (human immunodeficiency virus) (Human Immunodeficiency Virus), the purpose of treatment of AIDS." HAART " become the anti-AIDS prescription of standard at present.But " HAART " also no all roses, and it exists significant limitation.For example " HAART " curative effect is not 100%, patient must take medicine for a long time, and can not obtain radical cure, and also can cause toxic and side effects such as comprising lipodystrophy, back of the body stomach fat abnormal stacking, the unusual rising of serum lipid concentrations, wherein the most serious is liver failure and diabetes.And the expense of HAATR costliness also limited its use, surpasses 90% patient in the world and can not use it to treat.Reality is to press for to develop the inexpensive alternative medicine of treatment acquired immune deficiency syndrome (AIDS).Under such situation, traditional medicine causes people's attention unusually.There is the cultural tradition in more than 5,000 year in China, particularly original in style Chinese medicine is a wonderful work in the World Science culture, owing to have unique theoretical system and application method, ought in preventing and treating acquired immune deficiency syndrome (AIDS), bring into play bigger effect, for capturing acquired immune deficiency syndrome (AIDS), the world makes contributions.Though acquired immune deficiency syndrome (AIDS) is a kind of new human diseases, Chinese medicine document and experience be record to some extent never, learns based on traditional Chinese medical science symptom and etiology and pathology, and diagnosis and treatment acquired immune deficiency syndrome (AIDS) and relevant syndrome are fully feasible.Especially the traditional natural drug of several thousand flavors is gone through clinical practice in thousands of years, therefrom a medicine of preventing and treating acquired immune deficiency syndrome (AIDS) of developing suitable curative effect surely.Based on this, the research of natural inhibition HIV (human immunodeficiency virus) (Human Immunodeficiency Virus) (HIV) medicine is sought just in mushroom development in the whole world at present.
Hainan Island is an area that resources of medicinal plant is very abundant, and according to the record of Hainan flora, Hainan has floristics kind more than 4600 now, and kind surplus wherein medicinal plants just reaches 3100 accounts for 1/3 of the existing medicinal plants of China.Trigonostemon xyphophylloides is Euphorbiaceae (Euphorbiaceae) Trigonostemon (Trigonostemon genus) plant, is Hainan endemic species, mainly is distributed in Sanya and Lingshui.Trigonostemon xyphophylloides is among the people as anti-asthmatic in Hainan, find through consult the domestic and foreign literature data comprehensively, except that there being the scholar that Trigonostemon chinensis Trigonostemon reidioides was carried out not seeing the report about Trigonostemon xyphophylloides chemical constituent and pharmacology activity research chemical constituent and the pharmacology activity research abroad.
Summary of the invention
The purpose of this invention is to provide a kind of application of Trigonostemon xyphophylloides extract, it has significant anti-hiv activity.
The technical solution used in the present invention is: a kind of application of Trigonostemon xyphophylloides extract, the application of this extract in preparation AIDS virus resisting medicine.
The Trigonostemon xyphophylloides extract is about to the plant extract of Trigonostemon xyphophylloides through the organic solvent extraction gained.
The leaching process of described Trigonostemon xyphophylloides extract is: with Trigonostemon xyphophylloides oven dry and be ground into powder, and vacuum drying is to constant weight, with organic solvent reflux, extract, three times, merge extractive liquid,, filter, reclaim under reduced pressure is ethanol to the greatest extent, and the lyophilization powdered is the Trigonostemon xyphophylloides extract.Described organic solvent is ethanol or methanol.
Detect by the activity of application modern pharmacology experimental technique, find that this extract has significant AIDS virus resisting effect, can be used as the medicine of preparation AIDS virus resisting sword-like leave three essentials-essence art extract.
The present invention has following advantage:
1, plant bioavailability height, effect is remarkable.The present invention adopts organic solvent to extract, the active component dissolution rate height of Trigonostemon xyphophylloides, and extract has significant AIDS virus resisting effect.
2, the raw material sources uniqueness is safe.Trigonostemon xyphophylloides mainly is distributed in ground such as the Sanya in China Hainan Province and Lingshui, be the peculiar medicinal plants in Hainan, thereby resource has uniqueness; Extract provided by the present invention comes from natural plants, its among the people also for a long time as anti-asthmatic, therefore as medicine to side effect such as human body nonhazardouss.
3, development prospect is wide.Utilize the extract of natural plants to open up the new way of a treatment acquired immune deficiency syndrome (AIDS) or viral disease, and the further active component in the separation and purification extract, design and develop out the medicine for the treatment of acquired immune deficiency syndrome (AIDS) or viral disease more efficiently, applied range.
Description of drawings
Fig. 1 is the influence that the Trigonostemon xyphophylloides extract duplicates HIV.
Among the figure: (HIV) the Jurkat cell does not have infected by HIV to Ctrl;
Ctrl (+HIV) the Jurkat cell infection extract-treated of no use behind the HIV;
The solvent of DMSO dissolving extract is as negative control;
The AZT inverase, positive control;
TXE Trigonostemon xyphophylloides ethanol extraction.
Fig. 2 is the influence of Trigonostemon xyphophylloides extract pair cell survival.
Among the figure: (HIV) the Jurkat cell does not have infected by HIV to Ctrl;
Ctrl (+HIV) the Jurkat cell infection extract-treated of no use behind the HIV;
The solvent of DMSO dissolving extract is as negative control;
The AZT inverase, positive control;
TXE Trigonostemon xyphophylloides ethanol extraction.
Fig. 3 is the influence of Trigonostemon xyphophylloides extract to Jurkat cell proliferation and survival.
Among the figure: Ctrl: cell does not carry out any processing;
The solvent of DMSO dissolving extract is as negative control;
The AZT inverase, positive control;
TXE Trigonostemon xyphophylloides ethanol extraction.
Fig. 4 is that the Jurkat cell that the Trigonostemon xyphophylloides extract infects HIV-1 closes the influence that inclusion forms.
Among the figure: Moc does not have the Jurkat cell of infection;
Do not carry out the Jurkat cell of any processing after the Non infected by HIV-1;
The solvent of DMSO dissolving extract is as negative control;
The AZT inverase, positive control;
TXE Trigonostemon xyphophylloides ethanol extraction.
Fig. 5 is the direct influence of Trigonostemon xyphophylloides extract to reverse transcriptase activity.
Among the figure: Moc does not have the Jurkat cell of infection;
The PBS phosphate buffered saline(PBS) is as solvent control;
The solvent of DMSO dissolving extract is as negative control;
The AZT inverase, positive control;
TXE Trigonostemon xyphophylloides ethanol extraction.
Fig. 6 is the Trigonostemon xyphophylloides extract enters the U87.CD4.CXCR4 cell to HIV-1 influence.
Among the figure: the solvent of DMSO dissolving extract, as negative control;
TXE Trigonostemon xyphophylloides ethanol extraction;
The ENV virus capsid protein;
HXB2 contains the HIV-Luc virus of HXB2 coat protein;
VSV-G contains the HIV-Luc virus of VSV-G coat protein.
Fig. 7 is the influence of Trigonostemon xyphophylloides extract to HIV-1 gene expression.
Among the figure: the solvent of DMSO dissolving extract, as negative control;
TXE Trigonostemon xyphophylloides ethanol extraction;
The ENV virus capsid protein;
HXB2 contains the HIV-Luc virus of HXB2 coat protein.
The specific embodiment
The invention will be further described with indefiniteness embodiment below.
Embodiment one,
The Trigonostemon xyphophylloides stem is ground into powder and vacuum drying to constant weight, extracts with 80% methanol eddy, merge extractive liquid, filters, and reclaim under reduced pressure is methanol to the greatest extent, and merge extractive liquid, filters, and reclaim under reduced pressure is ethanol to the greatest extent, and the lyophilization powdered is the Trigonostemon xyphophylloides extract.
Embodiment two,
Trigonostemon xyphophylloides is ground into powder and vacuum drying to constant weight, uses 75% alcohol reflux, merge extractive liquid, filters, and reclaim under reduced pressure is ethanol to the greatest extent, and merge extractive liquid, filters, and reclaim under reduced pressure is ethanol to the greatest extent, and the lyophilization powdered is the Trigonostemon xyphophylloides extract.
Utilize the active example of Trigonostemon xyphophylloides extract anti HIV-1 virus:
1. treat the reagent thing
The powder of Trigonostemon xyphophylloides extract is dissolved in the dimethyl sulfoxine (DMSO) by 100mg/ml, and at room temperature jolting is spent the night gently.Filter to remove any insoluble granule with 3000 rev/mins of filtering heads with 0.4 μ m after centrifugal 10 minutes.Limpid part is as treating the reagent thing.
2. cell, virus and medicine
Jurkat cell and U87.CD4.CXCR4 cell are cultivated respectively in RPM1640 and DMEM, replenish 10% newborn calf serum, 100units/ml penicillin and 100ug/ml streptomycin sulfate.The proviral DNA of T preferendum HIV-1 cell strain HXBc2 advances the Jurkat cell to produce virus inoculation by the transfection of DEAE-Dextran method.HIV-1 HXBc2 Strain has been expressed other HXB2 gene of a part of composition of Eli Strain of HIV-1 and all.
3. HIV-1 duplicates experiment
In order to monitor virus replication, 1 * 10
6/ ml Jurkat cell at first infects with HIV-1, is equivalent to 10,000cpm reverse transcriptase (RT) activity.After infecting 24 hours, the RPMI-1640 complete medium is washed the virus that does not enter cell off entirely, adds the DMSO solvent processing for the treatment of reagent thing or equivalent.Measure the RT activity of each cell culture fluid after 48 hours, add new medicine simultaneously.With the chemical compound azidothymidine AZT (AZT, 3 '-Azido-3 '-deoxythymidine) as positive control.
4. cytotoxicity experiment
Treat cytotoxicity trypan blue exclusion method (the trypan blueexclusion method) mensuration of reagent thing.Just infected or do not have infected Jurkat cell to be present in to treat after the time different in the reagent thing with 0.2% trypan blue solution-dyed, living cell counting under light microscopic by HIV-1.
5. the preparation of HIV-1 pseudovirus and infection
By the pHXB2-env of calcium phosphate precipitation method with 20 μ g HIV-Luc plasmids and 4 μ g, pVSV-G, or pcDNA3 transfection 293T cells.Supernatant at culture fluid change back cell culture fluid after 48 hours is collected, and filters and save as viral liquid storage.Be equivalent to the active pseudovirus of 2000cpmRT and be used to infect the U87.CD4.CXCR4 cell.After 2 hours infection, repeatedly wash with fresh culture and to have removed original virus.Cell continues to have cultivated to collect after 48 hours does the active detection of Luc.
6. data analysis
All numerical value all uses meansigma methods ± S.E.M to represent, perhaps at least three independent experiments of representative.Relatively carrying out between the group with two tail student experimentings.P＜0.05 usefulness (*) mark, p＜0.01 usefulness (* *) mark.
7. testing result:
The ethanol extraction of Trigonostemon xyphophylloides stem (TXE) suppresses duplicating of HIV-1 and not obviously influence (Fig. 1, Fig. 2 and Fig. 3) of on cell proliferation and survival.This conclusion has the cell that stops HIV to infect and forms and close inclusion and further confirmed (Fig. 4) by finding it.Yet we do not detect its influence (Fig. 5) for the HIV-1RT enzymatic activity.We find that TXE can stop HIV-1 target approach cell (Fig. 6) potentially, and it is to entering back HIV-1 gene expression of cells influence very little (Fig. 7).These results have disclosed Trigonostemon xyphophylloides stem ethanol extraction and have had HIV (human immunodeficiency virus)-resistant activity by stoping entering of HIV-1 cell.
Claims (2)
1, the application of a kind of Trigonostemon xyphophylloides extract in preparation AIDS virus resisting medicine is characterized in that: the application of this extract in preparation AIDS virus resisting medicine; The preparation process of described extract is with the plant extract of Trigonostemon xyphophylloides through the organic solvent extraction gained.
2, the application of Trigonostemon xyphophylloides extract according to claim 1 in preparation AIDS virus resisting medicine, it is characterized in that: described organic solvent is ethanol or methanol.
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| CNA2008102135031A CN101347492A (en) | 2008-09-02 | 2008-09-02 | Use of Trigonostemon xyphophylloides extract in preparing anti-AIDS medicament |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102724990A (en) * | 2009-07-29 | 2012-10-10 | 印第安纳大学研究及科技有限公司 | Extracts of medicinal plant and uses thereof |
| CN108440458A (en) * | 2018-04-10 | 2018-08-24 | 海南师范大学 | A kind of new tetrahydrofuran derivatives class compound and its preparation method and application |
-
2008
- 2008-09-02 CN CNA2008102135031A patent/CN101347492A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102724990A (en) * | 2009-07-29 | 2012-10-10 | 印第安纳大学研究及科技有限公司 | Extracts of medicinal plant and uses thereof |
| CN108440458A (en) * | 2018-04-10 | 2018-08-24 | 海南师范大学 | A kind of new tetrahydrofuran derivatives class compound and its preparation method and application |
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Open date: 20090121 |