CN1843340A - Use of rhubarb anthraquinone compound in preventing and treating respiratory syncytial virus infectious disease - Google Patents
Use of rhubarb anthraquinone compound in preventing and treating respiratory syncytial virus infectious disease Download PDFInfo
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- CN1843340A CN1843340A CN 200510020685 CN200510020685A CN1843340A CN 1843340 A CN1843340 A CN 1843340A CN 200510020685 CN200510020685 CN 200510020685 CN 200510020685 A CN200510020685 A CN 200510020685A CN 1843340 A CN1843340 A CN 1843340A
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- syncytial virus
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Abstract
The invention relates to an application of anthraquinones compound in preventing diseases infected by respiratory syncytial virus. The median poison concentration TC50 of anthraquinones compound for cell Hep-2 is 209.7 mg/ml; the median effective concentration LC50 for respiratory syncytial virus is 96.7 mg/ml; the treating index is 2.3; the inhibiting rate to cytopathic RSV cell is over 50% when drug concentration is over 128 ug/ml; and the larger of drug concentration, the higher of inhibiting rate. The side effect of nthraquinones compound is smaller than that of ribavirin, the process is simple and it can be prepared into various dosage forms.
Description
(1) technical field:
The present invention relates to the application of rhubarb anthraquinone compound, the application of rhubarb anthraquinone compound on the control respiratory syncytial virus infectious disease specifically.
(2) background technology:
Respiratory syncytial virus (RSV) belongs to the Paramyxoviridae pneumonitis virus and belongs to, be all a genus with parainfluenza virus element etc., be single negative burst RNA viruses, can cause and go up lower respiratory illness, be infant and the common cause of disease of children acute lower respiratory infection in age in off year, nearly all child below 2 years old had rsv infection
[8]Find again that in recent years rsv infection is old people and immunosuppressant adult's an important virus
[9]At present rsv infection is still lacked special effective Therapeutic Method.The antiviral drugs virazole (have another name called ribavirin, Ribavirin) be proved to be and can be used for the treatment of rsv infection, but because its toxicity is bigger, the suction that can only atomize, and, make its clinical practice limited in recent years along with dispute to its curative effect
[10]Still do not have at present the vaccine of anti-rsv infection safely and effectively, therefore the research of anti-RSV medicine becomes the problem that international in recent years medical educational circles attractes attention.
(3) summary of the invention:
The objective of the invention is to disclose the low Chinese herb rhubarb extract of toxic and side effects---rhubarb anthraquinone compound is in the application that prevents and treats respiratory syncytial virus perception disease.
The present invention is applied to prevent and treat respiratory syncytial virus infectious disease with rhubarb anthraquinone compound.
Described rhubarb anthraquinone compound is in the application of control on the respiratory syncytial virus infectious disease, is rhubarb anthraquinone compound is made any dosage form on the pharmaceutics.
Rhubarb anthraquinone compound is made rhubarb injection, and total Radix Et Rhizoma Rhei anthraquinone content is 100mg/ml, every 2.0ml, clinically be used to prevent and treat the disease that causes by respiratory syncytial virus, usage: each 2.0ml, intramuscular injection, intravenous injection or intravenous drip, every day 2 times, 5 days courses of treatment.
Also rhubarb anthraquinone compound can be made the rhubarb tablets tablet, total Radix Et Rhizoma Rhei anthraquinone content is the 200mg/0.5g/ sheet, clinically is used to prevent and treat the disease that is caused by respiratory syncytial virus.The each 2-4 sheet of usage, oral, every day 2-3 time, 7 days is a course of treatment.
The present invention with the rhubarb anthraquinone compound anti respiratory syncytial virus be with the rhubarb anthraquinone compound of variable concentrations respectively interaction in vitro in the Hep-2 cell be seeded in respiratory syncytial virus on the Hep-2 cell, with the virazole is the contrast medicine, observe rhubarb anthraquinone compound to the toxic action of Hep-2 cell with to the inhibiting situation of respiratory syncytial virus, it is to the half toxic concentration TC of Hep-2 cell
50Be 209.7 μ g/ml, contrast is 364.7 μ g/ml, to the medium effective concentration IC of respiratory syncytial virus
50Be 96.7 μ g/ml, contrast is 66 μ g/ml.
Application result confirms that rhubarb anthraquinone compound is external has the obvious suppression effect to the respiratory syncytial virus that is seeded on the Hep-2 cell, and there is tangible dose-effect relationship, increase with drug level, its antiviral increased activity, suppression ratio raises, through linear regression analysis, and P＜0.05, rhubarb anthraquinone compound concentration is when 128 μ g/ml are above, and the cytopathic suppression ratio of RSV that respiratory syncytial virus is caused can reach more than 50%.
The concrete steps of using are:
1, used medicine is rhubarb anthraquinone compound (RAA), and cell is the Hep-2 cell, and virus is respiratory syncytial virus RSV-long strain;
2, cultivate the Hep-2 cell with cell culture technology, well-grown Hep-2 cell is dispersed into the individual cells suspension, by 1 * 10 with pancreatin
5/ ml concentration branch is planted in 40 orifice plates, every hole 0.1ml.Put 37 ℃, 5%CO
2Cultivate 24h in the incubator.It is standby to treat that cell grows up to behind the monolayer;
3, observe the toxic action of medicine to the Hep-2 cell:
Tetramethyl azo azoles salt (MTT) the method detection of drugs of setting up according to Mosmnna is to the toxic action of Hep-2 cell.
Grow up to the cell orifice plate of monolayer in cultivation, abandon the culture fluid supernatant, change the pastille that contains variable concentrations and keep liquid, every kind of concentration repeats 4 holes, and establishes the normal cell contrast.After continuing to cultivate 48h, abandon the culture fluid supernatant, every hole adds the MEM culture medium that does not the contain serum 50 μ l that contain 5mg/ml MTT, puts CO
2After continuing in the incubator to cultivate 2～3h, abandon the MTT supernatant, PBS washes 3 times, every hole adds lysate (DMSO: the ethanol volume ratio is 1: 1) 100 μ l, vibrates 5～10 minutes, dissolving fully to be crystallized, put on the 96 hole enzyme mark analyzers, measure the optical density OD value at 570nm wavelength place, and calculate cell survival rate, find out the maximal non-toxic concentration range of medicine pair cell according to following formula.
Cell survival rate (%)=(experimental port OD value/control wells OD value) * 100%.
Medicine shows as the Hep-2 cytotoxic effect: cell rounding, and wall thickness, the endochylema endoparticle increases, and refractivity strengthens.The drug effect of variable concentrations be the results are shown in Table 1 behind cell:
Table 1 rhubarb anthraquinone compound (RAA) is to the toxic action of Hep-2 cell
Table 1 The Cytotoxity of the RAA to Hep-2 cell
| Medicine compounds | Cell survival rate (%) The survival rate of cell (%) at various concentrations (μ g/ml) of the compounds during different pharmaceutical concentration (μ g/ml) | 50% toxic concentration TC 50 | ||||||||
| 10 | 20 | 40 | 80 | 160 | 320 | 640 | 800 | 1600 | ||
| RAA Ribavirin | 90.5 100.0 | 72.2 80.2 | 60.5 76.5 | 50.8 65.0 | 37.5 52.5 | 30.4 34.2 | 20.1 25.1 | 11.4 13.2 | 5.5 8.5 | 209.7 364.7 |
4, observe medicine to the effect of respiratory syncytial virus inhibition of proliferation:
On 40 orifice plates that grow up to monolayer Hep-2 cell, every hole inoculation 50ul 100TCID
50RSV liquid, respectively at 33 ℃ absorption 90min, abandon viral supernatant.According to the result of cytotoxicity experiment, in medicine non-toxic concn scope, the pastille that adds variable concentrations is kept the every hole 0.2ml of liquid, then the RSV culture plate is put 33 ℃, 5%CO
2Cultivate, observe CPE every day.Renew bright pastille every other day and keep liquid.Normal cell matched group, virus control group, positive drug virazole group and medicine group are established in experiment simultaneously.
The CPE recording method is :-be no CPE ,+be that CPE appears in 25% cell; ++ be that CPE appears in 50% cell; CPE appears in +++be 75% cell; ++ ++ be that CPE appears in 100% cell.About virus control ++ +～++ ++ the time abandon the culture fluid supernatant, every hole adds the culture fluid 50 μ l that contain 5mg/ml MTT, continues to cultivate flush away MTT supernatant behind 2～3h, add the every hole 100 μ l of DMSO lysate, mixing after 5～10 minutes, is surveyed the optical density OD value at 570nm wavelength place with microplate reader.And press following formula and calculate medicine the RSV suppression ratio.
RSV shows as the cellular swelling, becomes circle Hep-2 cell CPE, and refractivity weakens, and has syncytium to form.According to the result of cytotoxicity experiment, be chosen in the non-toxic concn scope, medicine is suppressed the experiment of RSV virus with different concentration respectively, its viral suppression ratio sees Table 2.
The anti-RSV effect of table 2 rhubarb anthraquinone compound (RAA)
Table 2 The antiviral activity of Rhubarb Anthraquinone analogues to RSV
| Medicine compounds | Different pharmaceutical concentration (viral suppression ratio (%) The inhibitory rate of RSV (%) at various concentrations (μ g/ml) of the compounds during g μ/ml) | 50% valid density IC 50 | Therapeutic index TI | ||||||
| 2 | 4 | 8 | 16 | 32 | 64 | 128 | |||
| RAA Ribavirin | 2.9 3.2 | 6.2 5.1 | 18.0 10.5 | 20.5 11.1 | 34.6 13.5 | 40.2 36.2 | 58.2 100.0 | 96.7 66.0 | 2.3 5.5 |
5,, calculate the half toxic concentration TC of medicine to the Hep-2 cell with the probit Return Law of the SPSS10.0 of statistical software in conjunction with the cytotoxicity experiment result
50Be 209.7 μ g/ml, the medium effective concentration IC of anti respiratory syncytial virus
50Be 96.7 μ g/ml, the therapeutic index Tl of medicine is 2.3, illustrates that rhubarb anthraquinone compound has stronger inhibitory action to respiratory syncytial virus in the Hep-2 cell.
Rhubarb anthraquinone compound is that the Chinese herb rhubarb extraction separation from China obtains, Radix Et Rhizoma Rhei is long at China's applicating history, ABUNDANT NATUREAL RESOURSES, the raw material Radix Et Rhizoma Rhei is easy to get, the effect of rhubarb anthraquinone compound anti respiratory syncytial virus is strong, and is little to the toxic and side effects of human body, the processing extraction technique maturation, can make injection, tablets and other formulations, be applied in the control of respiratory syncytial virus infectious disease.
(4) specific embodiment:
Embodiment 1: rhubarb anthraquinone compound is made the injection rhubarb injection, total Radix Et Rhizoma Rhei anthraquinone content 100mg/ml, every 2.0ml, clinically be used to prevent and treat the disease that causes by respiratory syncytial virus, usage: each 2.0ml, intramuscular injection, intravenous injection or intravenous drip, every day 2 times, 5 days courses of treatment.
Embodiment 2: rhubarb anthraquinone compound is made the rhubarb tablets tablet, and total Radix Et Rhizoma Rhei anthraquinone content 200mg/0.5g/ sheet clinically is used to prevent and treat the disease that is caused by respiratory syncytial virus.The each 2-4 sheet of usage, oral, every day 2-3 time, 7 days is a course of treatment.
Claims (4)
1, rhubarb anthraquinone compound is characterized in that: rhubarb anthraquinone compound is applied to prevent and treat respiratory syncytial virus infectious disease in the application of control respiratory syncytial virus infectious disease.
2, rhubarb anthraquinone compound according to claim 1 is characterized in that in the application of control respiratory syncytial virus infectious disease: rhubarb anthraquinone compound is made any dosage form on the pharmaceutics.
3, rhubarb anthraquinone compound according to claim 1 and 2 is in the application of control respiratory syncytial virus infectious disease, it is characterized in that: rhubarb anthraquinone compound is made rhubarb injection, total Radix Et Rhizoma Rhei anthraquinone content 100mg/ml, every 2.0ml, the clinical disease that causes by respiratory syncytial virus that is used for the treatment of, usage: each 2.0ml, intramuscular injection, intravenous injection or intravenous drip, every day 2 times, 5 days courses of treatment.
4, rhubarb anthraquinone compound according to claim 1 and 2 is inhaled the application of road syncytial virus infectious disease in control, it is characterized in that: rhubarb anthraquinone compound is made the Radix Et Rhizoma Rhei tablet, total Radix Et Rhizoma Rhei anthraquinone content 200mg/0.5g/ sheet, clinically be used to prevent and treat the disease that causes by respiratory syncytial virus, the each 2-4 sheet of usage, oral, every day 2-3 time, 7 days is a course of treatment.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103048450A (en) * | 2011-10-13 | 2013-04-17 | 上海药明康德新药开发有限公司 | Quantitative detection method of high-flux RSV (Respiratory Syncytial Virus) protein content and detection kit thereof |
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2005
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103048450A (en) * | 2011-10-13 | 2013-04-17 | 上海药明康德新药开发有限公司 | Quantitative detection method of high-flux RSV (Respiratory Syncytial Virus) protein content and detection kit thereof |
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