CN1631422A - Application of violet gold extractive in preparing process of medicine for resisting virus and bacteria - Google Patents
Application of violet gold extractive in preparing process of medicine for resisting virus and bacteria Download PDFInfo
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- CN1631422A CN1631422A CN 200410081402 CN200410081402A CN1631422A CN 1631422 A CN1631422 A CN 1631422A CN 200410081402 CN200410081402 CN 200410081402 CN 200410081402 A CN200410081402 A CN 200410081402A CN 1631422 A CN1631422 A CN 1631422A
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- radix ceratostigmatis
- ceratostigmatis mini
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Abstract
The invention discloses the application of violet gold extractive in preparing process of medicine for resisting virus and bacteria, which includes inhibition of virogene reproduction, transcription and translation, and in preparing antiviral drugs. It has strong bacterium resisting and antibiotic actions, and especially has strong inhibition and eradication actions to Staphylococcus aureus, Staphylococcus aureus drug resistant strain, and Haemophilus influenzae.
Description
Technical field:
The present invention relates to a kind of purposes of pure Chinese medicinal plant extract, especially relate to the purposes of Radix Ceratostigmatis mini plant extract aspect preparation antiviral and antibacterials.
Background technology:
Radix Ceratostigmatis mini (ceratostigma minus) is distributed in China southwest for the lumbaginaceae Radix Ceratostigmatis mini belongs to undershrub, and is wherein abundant with Tibet Autonomous Region's resource again.
According to " research of phenolic compound in the frame canopy "---" Botany Gazette " 1998,40 (11): 1035-1039 and " frame canopy The Chemical Constituents "---" Yunnan plant research " 1994, the record of 16 pertinent literatures such as (1:81-84), the separation of Radix Ceratostigmatis mini chemical analysis, structure identify that the main method that adopts is as follows: after the dry powder of Radix Ceratostigmatis mini is obtained extractum with 95% ethanol lixiviate, use petroleum ether, ethanol ethyl ester, n-butyl alcohol, hexane extraction respectively, by certain chromatography solvent burden ratio, a series of new separation material (as RP-
18, MCI-gel CHP
20P etc.) and modern spectroscopic technique (as
1HNMR,
13CNMR etc.) and chemical method, obtain 23 chemical compounds altogether, they are plumbasidesA-C, plumbocatechinsA and B, maltol glycoside plumbolactoneA, maltol-0-glucoside, plumbagin plumbagin, Quercetin quercetin, myricetin myricetin, Isoquercitrin quercetin3-0-glucoside, arbutin myricetin-3-0-rhamnoside, plumbagic acid plumbagic acid, isoshinanolone, epiisoshinanolon, N-trans-caf feoytyramine, N-trans-furuloyltyramine, acetovanilone apocynin, vanillic acid vanillic acid, syringic acid syringic acid, gallo catechin gallocatechin, (+)-cachou extract (+)-atechin and 1,2,6-three-O-galloyl glucose 1,2,6-tri-o-galloylglucoside, wherein plumbagin plumbagin and plumbagic acid plumbagic acid are Main Ingredients and Appearances.
Though being used as medicine, Radix Ceratostigmatis mini long history just arranged among the people, except that on the books in Tibetan medicine's masterpiece " brilliant pearl book on Chinese herbal medicine " of the 17th century in Christian era, in " Chinese medicine voluminous dictionary " (1988), " Chinese medicine voluminous dictionary " (1979), " choosing of Yunnan Chinese herbal medicine " (1970), " Tibetan medicine will " (1996), " Chinese herbal medicine is used in Tibet always " (1971), description to its purposes is also arranged in " national Chinese herbal medicine name mirror " books such as (1996), that is: Radix Ceratostigmatis mini has dredge the meridian passage, wind-damp dispelling, anti-inflammatory analgetic, the effect of reunion of fractured tendons and bones is mainly used in the treatment rheumatism, traumatic injury, fracture, vasculitis, parotitis, chronic lambago and skelalgia, menoxenia, headache and dizzy etc.Simultaneously, by Tibet Autonomous Region's key research project (1998-71) antiinflammatory, analgesia, the anastalsis Study of model of Radix Ceratostigmatis mini extractum are found: Radix Ceratostigmatis mini extractum has effect that suppresses mouse ear edema, the reaction of inhibition mice hot plate and the effect of shortening mice docking bleeding time and clotting time, thereby has further proved the effect of Radix Ceratostigmatis mini at aspects such as wind-damp dispelling, anti-inflammatory analgetic, hemostasis.But Radix Ceratostigmatis mini has the effect of too many levels such as suppressing that viral gene duplicates, transcribes and translates and had not but seen domestic and international report.
Though the antiviral drugs of clinical practice at present is more, but mostly be nucleoside derivate greatly, as ACV etc., its action target spot is a viral dna polymerase, and they directly reduce or the blocking virus dna polymerase activity with the form and normal substrate competition of enzyme reaction substrate analog, its common shortcoming is easily to produce the drug resistance strain, antiviral spectrum is narrow, and toxic and side effects is big, and price is high or pharmacological property is undesirable.
Summary of the invention:
The object of the present invention is to provide the purposes of Radix Ceratostigmatis mini plant extract aspect preparation antiviral and antibacterials.The present invention has brought into play new medical application to the Chinese medicine Radix Ceratostigmatis mini, opened up a new application, compare with existing synthetic drug and not only to have low toxicity advantage efficiently, but also have antiviral and antimicrobial dual function concurrently, it is safe and reliable, and drug action is strong, and good prospect in medicine is arranged, and Radix Ceratostigmatis mini abundant raw material, inexpensive, extraction process be simple, it is low to make the Chinese patent medicine cost, and can make various dosage forms.
For achieving the above object, the technical solution used in the present invention is as follows:
The invention is characterized in: the purposes of Radix Ceratostigmatis mini plant extract aspect preparation antiviral and antibacterials.
Described Radix Ceratostigmatis mini plant extract is used with single or compound recipe mode in preparation antiviral and antibacterials.
Described Radix Ceratostigmatis mini plant extract in the mode of its too many levels such as suppress that viral gene duplicates, transcribes and translates as the application in preparing antiviral drugs.
The application of described Radix Ceratostigmatis mini plant extract in preparation antiviral and antibiotic double effects medicine.
Described Radix Ceratostigmatis mini extract is with soft capsule or hard capsule or drop pill or tablet or dispersible tablet or granule or injection or oral liquid or suppository oral administration or skin or mucosa or rectal administration.
The invention has the advantages that:
1, Zijin is designated as pure Chinese medicine, and its safety of being used as medicine is very high;
2, the effect that has obvious anti-HSV-1 (herpes simplex virus I-type);
3, antibacterial, antibacterial action is strong, especially staphylococcus aureus, staphylococcus aureus persister, hemophilus influenza etc. is had good inhibition and killing action;
4, have antiviral and antimicrobial dual function, good prospect in medicine is arranged;
5, abundant raw material, inexpensive, extraction process are simple, to make the Chinese patent medicine cost low, and can make various dosage forms.
The specific embodiment:
Embodiment 1: inorganic and useful component content analysis
With the Radix Ceratostigmatis mini sample after the pulverization process, test its aminoacid kind and content value (W%) with the L-8800 of Hitachi type automatic amino acid analyzer; With reference to the related standards of health ministry fat, total sugar, mineral (ferrum, lead, magnesium, calcium, zinc, hydrargyrum, copper, the arsenic) content of Radix Ceratostigmatis mini sample are measured simultaneously.
The result shows, examined and contained 18 kinds of common amino acids in the Radix Ceratostigmatis mini sample altogether, wherein the highest with glycine content, reach 1.25E+00 (unit: W%), contain total sugar 28.3%, fat 1.52%, calcium 221.4mg/kg, magnesium 62.0mg/kg, zinc 0.70mg/kg, copper 0.50mg/kg, ferrum 5.42mg/kg, and the content of oxious component is very low, lead＜0.10mg/kg wherein, arsenic＜0.53mg/kg, hydrargyrum＜0.01mg/kg, the lead of stipulating in country health care (function) food universal standard GB16740-1997, arsenic, the content of hydrargyrum in solid product is no more than 2.0mg/kg respectively, 1.0mg/kg, 0.3mg/kg standard.Show that the safety that Radix Ceratostigmatis mini is used as medicine is very high.
Embodiment 2: acute toxicity test in mice
Get 20 mices, irritate stomach 150g/kg (being equivalent to original pharmaceutical content) with Radix Ceratostigmatis mini extractum once a day, observe the poisoning situation of animal in the administration 7 days.The result shows: animal occurs after 15 minutes in administration that action is slowed down, drowsiness phenomenon, recovers normal in about 4 hours.Death does not appear in the duration of test animal.Dissect also Non Apparent Abnormality of animal perusal during off-test, recording mice once irritates stomach Radix Ceratostigmatis mini ethanol extract maximum tolerated dose and is equivalent to crude drug in whole 150g/kg, this dosage is equivalent to the clinical application amount (by everyone used crude drug in whole gram number, everyone 50kg meter) more than 600 times, show that its toxicity is less.
Embodiment 3: antibiotic pharmacodynamics development test
Adopt the MH broth bouillon, with Radix Ceratostigmatis mini extractum do serial dilution become 1: 2～1: 128 concentration after, join respectively again in the culture medium of corresponding each bacterial strain, have or not bacterial growth in the observation culture medium, can determine minimal inhibitory concentration (MIC).Then, the liquid medium of no bacterial growth, transferred species has or not bacterial growth to plating medium in the observation plating medium, can determine minimal bactericidal concentration (MBC).
The result shows: Radix Ceratostigmatis mini extractum has shown stronger effect at antibiosis, staphylococcus aureus, staphylococcus aureus persister, hemophilus influenza etc. are had good inhibition and killing action, and its antibacterial MIC, MBC reach 1: 32 respectively and 1: 16.
Embodiment 4: the development test of antiviral drug effect
1, external pharmacodynamics test
Attacking the Vero cell with HSV-1 is cell model, adopts CPE inhibition method and mtt assay to study the drug influence of the external anti-HSV-1 of Radix Ceratostigmatis mini fluid extract.Experimental results show that Radix Ceratostigmatis mini and ACV therapeutic index there was no significant difference, find with transmission electron microscope observing simultaneously: use 625mgL HSV-1
-1The Radix Ceratostigmatis mini administration after virus obviously suppressed.In vitro tests shows: Radix Ceratostigmatis mini has anti-HSV-1 effect when being controlled at the non-toxic scope, and the big more effect of dosage is obvious more, and has direct kill virus granule, blocking virus to invade cell and the viral effect in time multiplexed cell system of inhibition.
2, pharmacodynamics test in the body
Make the pathogenic model of two kinds of HSV-1 by the local inoculation of the cornea that mice is carried out HSV-1, the inoculation of tail intravenous systemic.
A, herpes simplex keratitis model:
Draw " # " word with No. 4 syringe needles at mice cornea of right eye epithelium, drip HSV-1 viral suspension (TCID
50=10
5) 1 droplet, the massage 30s that closes one's eyes, the left eye contrast is dripped medicine every day 3 times.Behind the inoculation 36h, drip 2% fluorescein, observe the keratopathy situation with magnifier.Write down the keratopathy area every day.
B, mouse tail vein inoculation HSV-1 infection model:
70 mices are divided into seven groups, 10 every group.Test group is behind tail vein injection HSV-1, and second day beginning subcutaneous injection Radix Ceratostigmatis mini medicinal liquid treated 10 days altogether; Respectively at 1 of every group of execution in postvaccinal the 2nd, 4,8,12,16,20,30 day, under aseptic condition, get liver, kidney, spleen, heart and cerebral tissue.An organs and tissues part is put into 10% formalin solution, is used for doing routine pathology tissue slice (HE dyeing).The Radix Ceratostigmatis mini of non-toxic can alleviate the pathological change of internal organs in various degree, and high dose group (0.3g/kg/d) obviously reduces or eliminates the virus antigen in the internal organs.Experiment shows: Radix Ceratostigmatis mini can alleviate the pathological change of internal organs, obviously reduces or eliminates the virus antigen in the internal organs.
Found through experiments, the dosage of Radix Ceratostigmatis mini diluent eye drip is that 0.2mg//day seasonal disease variable area reduces slowly; Dosage is that 0.4mg//day pathological changes area reduces very fast; With treating over maximal non-toxic dosage 0.8mg//day, the pathological changes area reduces the fastest, and its curative effect is only put drops in one's eyes inferior slightly than ACV.Illustrate that Radix Ceratostigmatis mini is effective equally to the HSV-1 local infection, herpetic keratitis is had therapeutical effect.Illustrate that thus Radix Ceratostigmatis mini has tangible antagonism to viral HSV-1, its effect and traditional antiviral drugs acycloguanosine 1mg.ml
-1Similar.Though Zijin is designated as toxic herb, when the dosage of its mouse subcutaneous injection was low to moderate 0.3g/kg/d, clinical and pathology aspect did not all occur unusual, points out Radix Ceratostigmatis mini to have certain development prospect in the antiviral field thus.
Embodiment 5: pharmacology test
Montgomery in 1996
[10]Confirmed that Deng the people permissive cell surface specific receptor HVEM directly mediates HSV with HSV-1 glycoprotein gd specific bond and enters host cell.This test is by homemade dna probe corresponding to HSV-1 glycoprotein gd gene order, carried out dot blot hybridization and Northern hybridization, the Radix Ceratostigmatis mini of having found variable concentrations has been done further checking through the in situ hybridization of RT-PCR and cellular targets sheet to it simultaneously to the gene replication of HSV-1 glycoprotein gd and the influence of transcribing.
A, plaque method confirm: use 625mgL
-1The Radix Ceratostigmatis mini pretreatment cell can significantly suppress the absorption of HSV-1 to the Vero cell; Scanning electron microscope further confirms: use 625mgL
-1The Radix Ceratostigmatis mini pretreatment cell can significantly suppress the absorption of HSV-1 to the Vero cell.
B, self-control digoxin gD gene DNA probe carry out dot blot hybridization, and the result shows: gene replication has inhibitory action to Radix Ceratostigmatis mini to HSV-1gD; Different according to cell DNA with the HSV-1DNA buoyant density, further use the cesium chloride density gradient method with
3H-TdR mixes method and intuitively confirms: along with the increase of Radix Ceratostigmatis mini concentration, the HSV-1DNA peak progressively reduces, 40mgL
-1Radix Ceratostigmatis mini can 64.88% suppress the synthetic of HSV-1DNA, 625mgL
-1Radix Ceratostigmatis mini can 96.54% suppress the synthetic of HSV-1DNA, illustrate that thus Radix Ceratostigmatis mini has the obvious selectivity inhibitory action to HSV-1DNA synthetic.
The result of RT-PCR and Northern hybridization all shows: Radix Ceratostigmatis mini can suppress the expression of HSV-1 glycoprotein gd mRNA.The in situ hybridization result of cellular targets sheet shows the HSV-1 infected group of various dose Radix Ceratostigmatis mini effect, and the expression of its glycoprotein gd mRNA is lower than positive controls, and obviously increases with its suppression ratio of increase of drug level.HSV-1 behind counter immunoelectrophoresis, uses the HSV-1 behind the purification by the centrifugal purification of sucrose density gradient
3H-UTP mixes method and confirms: Radix Ceratostigmatis mini can obviously suppress the synthetic of the external RNA of HSV-1.
Claims (5)
1, the purposes of Radix Ceratostigmatis mini plant extract aspect preparation antiviral and antibacterials.
2, purposes according to claim 1 is characterized in that: the application of described Radix Ceratostigmatis mini plant extract in preparation antiviral and antibiotic double effects medicine.
3, purposes according to claim 1 and 2 is characterized in that: described Radix Ceratostigmatis mini plant extract is used with single or compound recipe mode in preparation antiviral and antibacterials.
4, purposes according to claim 1 and 2 is characterized in that: described Radix Ceratostigmatis mini plant extract in the mode of its too many levels such as suppress that viral gene duplicates, transcribes and translates as the application in preparing antiviral drugs.
5, purposes according to claim 1 and 2 is characterized in that: described Radix Ceratostigmatis mini extract is with soft capsule or hard capsule or drop pill or tablet or dispersible tablet or granule or injection or oral liquid or suppository oral administration or skin or mucosa or rectal administration.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410081402 CN1631422A (en) | 2004-12-07 | 2004-12-07 | Application of violet gold extractive in preparing process of medicine for resisting virus and bacteria |
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| CN 200410081402 CN1631422A (en) | 2004-12-07 | 2004-12-07 | Application of violet gold extractive in preparing process of medicine for resisting virus and bacteria |
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| CN1631422A true CN1631422A (en) | 2005-06-29 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101006827B (en) * | 2007-01-08 | 2010-10-06 | 冯耀宗 | Combination formula of green feed additive and its processing method and application |
| CN113951139A (en) * | 2021-11-08 | 2022-01-21 | 四川农业大学 | Creation method of novel anti-adversity cymbidium floribundum |
-
2004
- 2004-12-07 CN CN 200410081402 patent/CN1631422A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101006827B (en) * | 2007-01-08 | 2010-10-06 | 冯耀宗 | Combination formula of green feed additive and its processing method and application |
| CN113951139A (en) * | 2021-11-08 | 2022-01-21 | 四川农业大学 | Creation method of novel anti-adversity cymbidium floribundum |
| CN113951139B (en) * | 2021-11-08 | 2023-04-07 | 四川农业大学 | Creation method of novel anti-adversity cymbidium floribundum |
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