CN103520201A - Chinese medicinal polysaccharide compound with function of resisting new-castle disease viruses - Google Patents
Chinese medicinal polysaccharide compound with function of resisting new-castle disease viruses Download PDFInfo
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- CN103520201A CN103520201A CN201310282665.1A CN201310282665A CN103520201A CN 103520201 A CN103520201 A CN 103520201A CN 201310282665 A CN201310282665 A CN 201310282665A CN 103520201 A CN103520201 A CN 103520201A
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- polysaccharide compound
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Abstract
The invention discloses a Chinese medicinal polysaccharide compound with a function of resisting new-castle disease viruses, and belongs to the technical field of antiviral veterinary medicine researches. The Chinese medicinal polysaccharide compound is prepared by extracting respectively to prepare rhizoma polygonati polysaccharide and sulfated radix codonopsis pilosulae polysaccharide, and forming the compound by using the polysaccharides in a ratio of 9:1. The Chinese medicinal polysaccharide compound has a significant inhibitory effect on the new-castle disease viruses, and can be developed into a new veterinary antiviral medicament.
Description
One, technical field
A kind of herbal polysaccharide compound recipe with anti-new castle disease virus effect of the present invention, belongs to antiviral herbal medicine studying technological domain.
Two, background technology
Zoonosis is the development of viral infectious serious threat aquaculture always especially, causes huge economic loss.This class disease of prevention and control is mainly abutted against kind of a vaccine prevention, but for various reasons, is still difficult to control completely its outburst, as newcastle disease, bird flu, swine fever etc.Once these diseases are fallen ill, there is no the Drug therapy of specially good effect.Though some cytokines have certain curative effect, because it is expensive, be difficult to apply in livestock and poultry breeding industry.Therefore, find novel antiviral medicine is safely and effectively the focus of veterinary applications research always.
Large quantity research shows, much Chinese medicine and effective ingredient thereof have certain antivirus action, but a little less than their effect, Chinese medicine especially, due to complicated component, difficult quality is controlled, and is abroad difficult to accept.Nearest studies confirm that, sulphation modification can further improve the antiviral activity of herbal polysaccharide, and the effect of sulfated polysaccharides compound recipe is significantly better than single component medicine.Further research is found, although sulfated polysaccharides compound recipe has good effect, higher by the cost of synthetic sulfated polysaccharides, safety reduces, and is unfavorable for applying on veterinary clinic.With part polysaccharide, substitute after sulfated polysaccharides, the effect of compound recipe does not subtract, but cost significantly reduces, and safety improves, and is more conducive to apply on veterinary clinic.
The present invention by the active Comparison and Screening of herbal polysaccharide, the sulphation modification of polysaccharide and specific activity compared with a series of researchs such as, prescription screening, proportion compatibility screenings, filter out a kind of safe, effective, quality controllable, anti-new castle disease virus herbal polysaccharide compound recipe of having application prospect at veterinary clinic.
Three, summary of the invention
Technical problem the present invention is directed to the problem that veterinary clinic lacks antiviral drugs, and a kind of safe, effective, anti virus herb compound polyose of having application prospect at veterinary clinic is provided.
A kind of herbal polysaccharide compound recipe with anti-new castle disease virus effect of technical scheme, comprise: extract 5 kinds of polysaccharide, prepare 4 kinds of sulfated polysaccharides and 15 polysaccharide-sulfated polysaccharides compound recipes, measure their cytotoxicities and antiviral activity, filter out the compound recipe that antiviral activity is the strongest.It is characterized in that, polygonatum polysaccharide and sulphation Radix Codonopsis polysaccharide are the most remarkable to the inhibitory action of Avian pneumo-encephalitis virus by 9: 1 prescriptions with sugared content.
Beneficial effect the invention provides a kind of herbal polysaccharide compound recipe with anti-new castle disease virus effect, can be developed into as antivirus veterinary new drug, on animal produces, applies, and effectively improves the control effect of animal epidemic, ensures the sound development of aquaculture.
Compared with prior art, advantage of the present invention is as follows:
1. from need of production aspect, Field of Animal Epidemic Disease Control lacks effective, safe, economic Chinese and western drugs, the invention provides a kind of herbal polysaccharide compound recipe with anti-new castle disease virus effect, can be developed into antivirus veterinary new drug, fills up the blank of domestic and international veterinary drug research.
2. from herbal medicine research aspect, the present invention's application molecular modification technology is carried out structure of modification to herbal polysaccharide, and with polysaccharide-sulfated polysaccharides prescription, not only drug effect is reliable, and definite ingredients, quality controllable, has no report both at home and abroad.
3. compare with all sulfuric acid compound polyose, drug effect of the present invention does not reduce, but cost reduces (purification of 9 parts of polysaccharide and sulphation modification expense) greatly, and safety improves, is conducive to apply on veterinary clinic.
four, accompanying drawing explanation
Fig. 1 is the schematic diagram of the highest viral suppression ratio of each group.
Note: CP-sCA, Radix Codonopsis polysaccharide adds sulphation Radix Angelicae Sinensis polysaccharide compound recipe; EP-sAP, epimedium brevicornum polysaccharide adds sulphation astragalus polysaccharides compound; EP-sCA, epimedium brevicornum polysaccharide adds sulphation Radix Angelicae Sinensis polysaccharide compound recipe; CA-sEP, Radix Angelicae Sinensis polysaccharide adds sEPS compound recipe; SR-sCP polygonatum polysaccharide adds sulphation Radix Codonopsis polysaccharide compound recipe; PS<sub TranNum="70">9</sub>-sPS<sub TranNum="71">1</sub>, the proportioning of polysaccharide and sulfated polysaccharides is 9:1; PS<sub TranNum="72">7</sub>-sPS<sub TranNum="73">3</sub>, the proportioning of polysaccharide and sulfated polysaccharides is 7:3; PS<sub TranNum="74">6</sub>-sPS<sub TranNum="75">4</sub>, the proportioning of polysaccharide and sulfated polysaccharides is 6:4.On a-h post, mark alphabetical different person's significant differences (P < 0.05).
five, the specific embodiment
1. the preparation of polysaccharide extraction and sulfated polysaccharides
Radix Angelicae Sinensis polysaccharide (CA), astragalus polysaccharides (AP), epimedium brevicornum polysaccharide (EP), Radix Codonopsis polysaccharide (CP) and polygonatum polysaccharide (SR), extract with decoction alcohol precipitation method respectively; Sulphation Radix Angelicae Sinensis polysaccharide (sCA), sulphation astragalus polysaccharides (sAP), sEPS (sEP) and sulphation Radix Codonopsis polysaccharide (sCP), use respectively chlorosulfonic acid-pyridine legal system standby, with sulfuric acid-phynol method, measure polyoses content, barium chloride-gelatin method is measured degree of substitution.
2. the preparation of polysaccharide-sulfated polysaccharides compound recipe
Five kinds of prescriptions add sulphation Radix Angelicae Sinensis polysaccharide (CP-sCA), epimedium brevicornum polysaccharide by Radix Codonopsis polysaccharide respectively and add sulphation astragalus polysaccharides (EP-sAP), epimedium brevicornum polysaccharide and add that sulphation Radix Angelicae Sinensis polysaccharide (EP-sCA), Radix Angelicae Sinensis polysaccharide add sEPS (CA-sEP), polygonatum polysaccharide adds sulphation Radix Codonopsis polysaccharide (SR-sCP) and forms, and each prescription is 9: 1 (PS according to polysaccharide (PS) with the sugared content ratio of sulfated polysaccharides (sPS) respectively again
9-sPS
1), 7: 3 (PS
7-sPS
3), 6: 4 (PS
6-sPS
4) be made into 3 compound recipes, totally 15 prescriptions.
3. the cytotoxic assay of compound polyose
Use respectively MEM culture fluid from 500 μ gmL 15 prescriptions
-1doubling dilution to 0.977 μ gmL
-1totally 10 concentration.Get 9 age in days SPF Embryo Gallus domesticus (being purchased from Nanjing medical instruments factory) and prepare chick embryo fibroblast (CEF).Cell concentration is adjusted to 5 * 10
5mL
-1, being inoculated in 96 porocyte culture plates, every hole 100 μ L, put 38.5 ℃, 5%CO
2incubator is cultivated 24h, adds the prescription of each concentration, and every concentration repeats 4 holes, separately establishes cell control well, 37 ℃, 5%CO
2under condition, cultivate 72h, with mtt assay, measure cell A
570value.With A
570value is not significantly less than cell control well A
570the maximum concentration of value is as the maximum safe concentration of this prescription.As a result, the maximum safe concentration of each prescription is at 31.25～7.813 μ gmL
-1.For ease of relatively unified, each prescription maximum safe concentration unification is set as to 7.813 μ gmL
-1.
4. the comparison of the antiviral activity of compound polyose
By each compound recipe respectively from 7.813 μ gmL
-1doubling dilution to 0.488 μ gmL
-1totally 5 concentration.Cultivate CEF, when cell grows up to monolayer, every hole adds 100TCID
50newcastle disease virus (La Sota strain IV) liquid 100 μ L, put CO
2incubator is hatched 2h, removes virus liquid, adds respectively the prescription of each concentration.Every concentration repeats 4 holes, establishes virus control group (VC only adds virus), cell matched group (CC only adds MM culture fluid) and blank group (BC does not add cell), 37 ℃, 5%CO simultaneously
2under condition, cultivate 72h, with mtt assay, measure cell A
570value, as the index (A of each prescription antivirus action
570be worth greatlyr, antivirus action is stronger); And the highest viral suppression ratio of calculating
the power of each prescription antivirus action relatively.Result is as follows:
(1) PS
9-sPS
1the antiviral activity of each group
Each compound recipe group is at 7.813～0.488 μ gmL
-1cell A
570value is all significantly higher than virus control group (P < 0.05) (table 1), shows that they all have significant antivirus action in these concentration.
Table 1 PS
9-sPS
1the cell A of each group
570value
(2) antiviral activity of each group of PS7-sPS3
CP
7-sCA
3, EP
7-sAP
3, EP
7-sCA
3and CA
7-sEP
3all concentration groups, SR
7-sCP
3cell A at 3.906～0.488 groups
570value is significantly higher than virus control group (P < 0.05) (table 2).Show that they all have significant antivirus action in these concentration.
Table 2 PS
7-sPS
3the cell A of each group
570value
Note: same column data mark alphabetical different person's significant differences (P < 0.05).Following table is same.
(3) antiviral activity of each group of PS6-sPS4
CP
6-sCA
4, EP
6-sAP
4and SR
6-sCP
4all concentration groups, EP
6-sCA
4at 7.813 μ gmL
-1, CA
6-sEP
4at 3.906,0.977 μ gmL
-1the cell A of group
570value is significantly higher than virus control group (P < 0.05) (table 3).Show that they all have significant antivirus action in these concentration.
Each PS of table 3
6-sPS
4the cell A of compound recipe group
570value
(4) the highest viral suppression ratio of each group
The viral suppression ratio of three kinds of proportionings of same prescription is compared, CP-sCA, EP-sAP and EP-sCA are the highest at the viral suppression ratio of 9: 1 in 7: 3, CA-sEP and SR-sCP, are significantly higher than other two kinds of proportionings (P < 0.05) (accompanying drawing 1).Show that these proportionings are respectively their best proportionings.
The viral suppression ratio of 15 compound recipes is compared, SR
9-sCP
1the viral suppression ratio the highest (66.12%) of group, is significantly higher than all the other 14 groups (P < 0.05); Secondly be CP
7-sCA
3group (52.89%) is significantly higher than except SR
9-sCP
113 groups of all the other outside group; Secondly be EP again
7-sAP
3(42.18%), CA
9-sEP
1(41.20%), EP
7-sCA
3(39.0%) group (accompanying drawing 1).Show SR
9-sCP
1antiviral activity the strongest, can be used as the prescription of anti-new castle disease virus.
Claims (2)
1. a herbal polysaccharide compound recipe with anti-new castle disease virus effect, is characterized in that, with sugared content meter, and 9 parts of polygonatum polysaccharides, 1 part of sulphation Radix Codonopsis polysaccharide.
2. the herbal polysaccharide compound recipe that described in claim 1, method obtains, has significant inhibitory action to newcastle disease virus.
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| CN201310282665.1A CN103520201B (en) | 2013-07-08 | 2013-07-08 | A kind of have the polygonatum polysaccharide of anti-new castle disease virus effect and the compositions of sulphation Radix Codonopsis polysaccharide |
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| CN201310282665.1A CN103520201B (en) | 2013-07-08 | 2013-07-08 | A kind of have the polygonatum polysaccharide of anti-new castle disease virus effect and the compositions of sulphation Radix Codonopsis polysaccharide |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090252848A1 (en) * | 2008-04-03 | 2009-10-08 | Sunanta Pongsamart | Immunomodulating polysaccharide gel from durian fruit-rind as additive in animal feed |
| CN102225103A (en) * | 2011-05-30 | 2011-10-26 | 河南黑马动物药业有限公司 | Formulation and production process of traditional Chinese medicinal injection solution for controlling Newcastle disease |
| CN102690366A (en) * | 2012-06-11 | 2012-09-26 | 殷中琼 | Preparation method of plant polysaccharide sulfate with immune enhanced activity and anti-virus effect, as well as application |
| CN103169773A (en) * | 2011-12-20 | 2013-06-26 | 洛阳惠中兽药有限公司 | Traditional Chinese medicine compound preparation for enhancing livestock immunity and preparation method |
-
2013
- 2013-07-08 CN CN201310282665.1A patent/CN103520201B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090252848A1 (en) * | 2008-04-03 | 2009-10-08 | Sunanta Pongsamart | Immunomodulating polysaccharide gel from durian fruit-rind as additive in animal feed |
| CN102225103A (en) * | 2011-05-30 | 2011-10-26 | 河南黑马动物药业有限公司 | Formulation and production process of traditional Chinese medicinal injection solution for controlling Newcastle disease |
| CN103169773A (en) * | 2011-12-20 | 2013-06-26 | 洛阳惠中兽药有限公司 | Traditional Chinese medicine compound preparation for enhancing livestock immunity and preparation method |
| CN102690366A (en) * | 2012-06-11 | 2012-09-26 | 殷中琼 | Preparation method of plant polysaccharide sulfate with immune enhanced activity and anti-virus effect, as well as application |
Non-Patent Citations (3)
| Title |
|---|
| 乔海博 等: "复方中药多糖对鸡新城疫疫苗免疫效果的影响", 《石河子大学学报》, vol. 30, no. 6, 31 December 2012 (2012-12-31) * |
| 王君敏 等: "8种硫酸化多糖对新城疫病毒感染鸡胚成纤维细胞能力的影响", 《南京农业大学学报》, vol. 34, no. 1, 30 January 2011 (2011-01-30) * |
| 颜新艳 等: "硫酸化多糖复方和中药成分复方的鸡新城疫疫苗免疫增强效果比较", 《江苏农业科学》, 15 June 2010 (2010-06-15) * |
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