CN107468682A - Application of the mangiferin in antiviral drugs is prepared - Google Patents
Application of the mangiferin in antiviral drugs is prepared Download PDFInfo
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- CN107468682A CN107468682A CN201710855423.5A CN201710855423A CN107468682A CN 107468682 A CN107468682 A CN 107468682A CN 201710855423 A CN201710855423 A CN 201710855423A CN 107468682 A CN107468682 A CN 107468682A
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- mangiferin
- svcv
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- channel catfish
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
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- Pharmacology & Pharmacy (AREA)
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- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
The invention provides application of the mangiferin in antiviral drugs is prepared, including mangiferin to prepare the application in treating or preventing SVCV medicine, including mangiferin is preparing the application in treating or preventing channel catfish virus medicine.By pre-processing FHM cells or CCO cells with certain density mangiferin, then with SVCV or channel catfish virus infection, free virus is washed away, is continuously added containing corresponding concentration mangiferin, the levels of replication of virus is detected after 24h.As a result show, after mangiferin is handled, both viral levels of replication poles significantly reduce cell, illustrate that mangiferin has the function that suppression SVCV and channel catfish virus etc. are aquatic viral.Cytotoxicity experiment shows that the mangiferin of institute's concentration does not have toxic side effect to FHM and CCO cells, can be as the therapeutic agent and prophylactic of SVCV disease and channel catfish virosis.
Description
It for the applying date is 2015-7-24 that the application, which is, application number：201510444307.5 denomination of invention：Mangiferin exists
Prepare the divisional application of the application in antiviral drugs.
Technical field
The present invention relates to the purposes of mangiferin, and anti-SVCV or spot are being prepared more particularly to mangiferin
Application in Cha Wei Channel-catfish virus drugs.
Background technology
SVCV (Spring Viraemia of Carp Virus, SVCV), is under the jurisdiction of Rhabdoviridae
(Rhabdoviridae), vesicular stomatitis virus category (Vesiculovirus), caused spring viremia be it is a kind of it is acute,
Hemorrhagic and the disease for having serious infectious, by International Office of Epizootics (OIE) be classified as must notifiable disease, the Ministry of Agriculture also will
Be classified as《The People's Republic of China (PRC) enters the territory animal one, two class infectious diseases, parasitic disease register》(2008) a kind of animal epidemic disease in
Disease.
Channel catfish virus (CCV) is under the jurisdiction of herpesviral mesh (Herpesvirales), fish herpetoviridae
(Alloherpesviridae), Channel-catfish fish herpetovirus category (Ictalurivims), there is the double-stranded DNA virus of cyst membrane.It is caused
Channel catfish virosis can cause the mortality of fry and fingerling in plant, be classified as by International Office of Epizootics (OIE) necessary
It is also classified as by the disease declared, the Ministry of Agriculture《The People's Republic of China (PRC) enters the territory animal one, two class infectious diseases, parasitic disease
Register》(2008) two class animal epidemics in.
Disease is endangered China's culture fishery huge as caused by above-mentioned virus, temporarily without effective medicine and treatment
Method, once outburst can cause huge loss to culture fishery.Find anti-SVCV and channel catfish
Virus medicine be particularly important with it is urgent.
Mangiferin (mangiferin) also known as mangiferin, asphonin, are a kind of polyphenol compounds, and molecular formula is
C19H18O11, relative molecular weight 422.Mangiferin is mainly from the leaf of Anacardiaceae plant mango and almond, fruit, bark, rough gentian
Section's plant Northeastern Radix Gentianae, mussot swertia herb, Plants of Polypodiaceae balt pyrrosia herb etc. are isolated, and also someone divides from vegetables in recent years
Separate out mangiferin.As what mangiferin was studied deepens continuously, eliminating the phlegm that its is relieving cough and asthma, anti-inflammatory and antalgic, hepatic cholagogic is immunized, resists
A variety of pharmacological activity such as lipid peroxidation, anticancer, anti-diabetic, antibacterial, which are constantly excavated, to be come out.However, there is not yet its have it is anti-
The report of the Aquatic animals virus such as SVCV and channel catfish virus, therefore whether inventor has to mangiferin
There is anti-SVCV and channel catfish virus to be studied, it is found that it can significantly inhibit both viruses
Replicate, it is safe and non-toxic to cell under valid density, therefore it is great as the sick cytotoxic drug development significance of water resistant.
The content of the invention
It is an object of the invention to provide mangiferin in treatment or prevention SVCV medicine is prepared
Using, including mangiferin suppresses the application in SVCV in vitro.
It is another object of the present invention to provide mangiferin to prepare treatment or prevention channel catfish virus medicine
In application, including mangiferin suppresses the application in channel catfish virus drugs in vitro.
Final object of the present invention is that providing mangiferin is preparing while treating or preventing anti-spring viremia
Application in virosis and channel catfish virus medicine.
In order to achieve the above object, the present invention takes following technical measures：
The present invention has carried out the Antiviral breeding of cellular level with mangiferin, is pre-processed respectively with certain density mangiferin
FHM cells and CCO cells, then with 0.1MOI SVCV and channel catfish virus infection, wash away free
Virus, continuously add containing corresponding concentration mangiferin, the levels of replication of virus is detected after 24h.As a result show, cell is through mangiferin
After processing, both viral levels of replication poles significantly reduce, and illustrate that mangiferin has and suppress SVCV and spot
The aquatic viral effects such as point Cha Wei Channel-catfish viruses.Cytotoxicity experiment shows the mangiferin of institute's concentration to FHM and CCO cells
There is no toxic side effect, can be as the therapeutic agent and prophylactic of SVCV disease and channel catfish virosis.
Compared with prior art, the present invention has advantages below：
1. present invention firstly discovers that mangiferin can be used for the preventing and treating of aquatic products field virus, to treat SVCV
Disease and channel catfish virosis provide new therapy approach and means.
2. mangiferin can suppress SVCV duplication, CCV duplication can also be suppressed, and be a kind of low to cytotoxic
The efficient antiviral drugs of poison.
3. utilizing modern common drug preparation means, tablet, capsule, particle can be made using mangiferin as active component
Any pharmaceutically acceptable formulation such as agent, oral liquid, sustained release preparation, controlled release preparation, nanometer formulation, injection.
Brief description of the drawings
Fig. 1 be employed in the embodiment of the present invention 1 various concentrations mangiferin to the toxicity test result of FHM cells.
Fig. 2 be employed in the embodiment of the present invention 2 various concentrations mangiferin to SVCV inhibition assay result.
Fig. 3 be employed in the embodiment of the present invention 1 various concentrations mangiferin to the toxicity test result of CCO cells.
Fig. 4 be employed in the embodiment of the present invention 3 various concentrations mangiferin to CCV inhibition assay result.
Embodiment
The application of mangiferin of the present invention is illustrated below by example.These embodiments are only said to the present invention
It is bright, rather than limit the invention.Technical scheme of the present invention, it is the conventional skill of this area if not otherwise specified
Art.
Embodiment 1：
Mangiferin is to cytotoxicity experiment
Drug toxicity, specific method are detected using mtt assay：Collect fat head carp cell (the Fat head of exponential phase
Minnow cell, FHM) cell or CCO cells (channel catfishes gonad cell), concentration of cell suspension is adjusted, is added into
Into 96 orifice plates, 200 μ L nutrient solutions are added per hole, cell density is filled in 1000-5000/ holes, edge hole with sterile PBS；5%
CO2, 28 DEG C are incubated the mangiferin that concentration gradient can be added to (about 6-8h) after cell attachment, and each gradient sets 6 multiple holes, after
It is continuous to be incubated 48h, observe cell state under inverted microscope；20 μ L MTT solution (5mg/ml, i.e. 0.5%MTT) are added per hole, after
Culture is terminated after continuous culture 4h, it is careful to exhaust nutrient solution in hole；150 μ L dimethyl sulfoxide (DMSO)s are added per hole, are placed on decolorization swinging table
Low-speed oscillation 10min, makes crystal fully dissolve, and the light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument OD490nm, calculates
Cell survival rate.
This experiment sets zeroing hole (culture medium, MTT, dimethyl sulfoxide (DMSO)) simultaneously, control wells (cell, respective concentration it is molten
Agent, nutrient solution, MTT, dimethyl sulfoxide (DMSO)).
As a result as shown in figure 1, used by experiment after each concentration mangiferin processing FHM cells 24h cell survival rate with it is right
Change according to group compared to no conspicuousness, show the mangiferin of institute's concentration does not have toxic action to FHM cells.
As a result as shown in figure 3, after each concentration mangiferin processing CCO cells 24h employed in experiment cell survival rate with
Control group changes compared to no conspicuousness, and show the mangiferin of institute's concentration does not have toxic action to CCO cells.
Embodiment 2：
Mangiferin is preparing the application in treating or preventing SVCV medicine, comprises the following steps：
FHM is inoculated with 12 orifice plates, after 70-80% individual layers are grown to, nutrient solution is abandoned in suction, add containing 100uM, 50uM,
25uM, 12.5uM mangiferin and PBS complete culture solution pre-process 6h, and nutrient solution is abandoned in suction, are inhaled with the 0.1MOI μ L of SVCV 100
Attached cell, 28 DEG C of incubation 1h, washes away free virus, continuously adds the complete medium containing corresponding concentration mangiferin and PBS,
28 DEG C, continue in 5%CO2 incubators to cultivate 24h (processing of each concentration sets three independent repeating groups).Collected after 24h
Cell extraction total serum IgE, fluorogenic quantitative detection SVCV-G gene expression doses after reverse transcription.Quantitatively detect the primer of SVCV-G genes
Sequence is TGCTGTGTTGCTTGCACTTATYT/TCAAACKAARGACCGCATTTCG.The common encoding nuclear proteins (N) of SVCV, phosphorus egg
5 structural proteins such as RNA polymerase (L- albumen) of (P), memebrane protein (M), glycoprotein (G) and dependenc RNA in vain, wherein by G bases
It is related with virus virulence because the glycoprotein of coding determines SVCV host range, frequently as the detection target of virus load.
As a result as shown in Fig. 2 compared with control group, when mangiferin concentration is 25uM the expression of SVCV-G genes significantly drop
Low (P ＜ 0.05), decline 31.38% compared with control group.When mangiferin concentration is 50uM and 100uM, gene expression amount extremely significantly drops
Low (P ＜ 0.01), 36.57% and 67.42% are lowered compared with control group respectively.Show that mangiferin can suppress SVCV duplication, and
With concentration dependent.
Embodiment 3：
Mangiferin is preparing the application in treating or preventing channel catfish virus medicine, comprises the following steps：
CCO cells are inoculated with 12 orifice plates, after 70-80% individual layers are grown to, nutrient solution is abandoned in suction, add containing 100uM,
50uM, 25uM, 12.5uM mangiferin and PBS complete culture solution pre-process 6h, and nutrient solution is abandoned in suction, with the 0.1MOI μ of CCV 100
L adherent cells, 28 DEG C of incubation 1h, wash away free virus, continuously add the complete culture containing corresponding concentration mangiferin and PBS
Base, 28 DEG C, continue in 5%CO2 incubators to cultivate 24h (processing of each concentration sets three independent repeating groups).Received after 24h
Collect cell extraction total serum IgE, fluorogenic quantitative detection CCV-ORF46 gene expression doses after reverse transcription.Quantitatively detect CCV-ORF46 bases
The primer sequence of cause is GTCTATTTTGGAGGCTGC/ATTCGGGTTATCGGTGGC.CCV-ORF46 encodes CCV main knot
Structure albumen, it is related with the invasion of virus, frequently as the detection target of virus load.
As a result as shown in figure 4, compared with control group, when mangiferin concentration is 50uM, CCV levels of replication significantly reduces
(P ＜ 0.05), 41.43% is lowered compared with control group, CCV levels of replication pole significantly reduces (P when mangiferin concentration is 100uM
＜ 0.01), lower 76.07% compared with control group.As a result the anti-CCV of cell can be significantly improved after showing mangiferin processing CCO cells
Effect, and there is concentration dependent.
By above-mentioned it is demonstrated experimentally that mangiferin significantly reduces the duplication of SVCV and channel catfish virus
Level, it is safe and non-toxic to FHM cells and CCO cells.Mangiferin can be used for prepare prevention and treatment spring viremia or/and
The medicine of channel catfish virosis.
Claims (4)
1. mangiferin is preparing the application in treating or preventing channel catfish virus medicine.
2. mangiferin suppresses the application in channel catfish virus drugs in vitro.
3. mangiferin is in preparing while treating or preventing anti-SVCV disease and channel catfish virus medicine
Application.
4. according to the application described in claim 1 or 2 or 3, described medicine is any medically acceptable formulation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710855423.5A CN107468682B (en) | 2015-07-24 | 2015-07-24 | Application of mangiferin in preparation of antiviral drugs |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710855423.5A CN107468682B (en) | 2015-07-24 | 2015-07-24 | Application of mangiferin in preparation of antiviral drugs |
| CN201510444307.5A CN104997769A (en) | 2015-07-24 | 2015-07-24 | Applications of mangiferin in preparation of antiviral drugs |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
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| CN201510444307.5A Division CN104997769A (en) | 2015-07-24 | 2015-07-24 | Applications of mangiferin in preparation of antiviral drugs |
Publications (2)
| Publication Number | Publication Date |
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| CN107468682A true CN107468682A (en) | 2017-12-15 |
| CN107468682B CN107468682B (en) | 2021-03-02 |
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| CN201710855423.5A Active CN107468682B (en) | 2015-07-24 | 2015-07-24 | Application of mangiferin in preparation of antiviral drugs |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114533799A (en) * | 2022-03-17 | 2022-05-27 | 湖南师范大学 | Application of callicarpa nudiflora in antiviral aspect of freshwater fishes |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111166737B (en) * | 2019-10-21 | 2022-04-01 | 江苏农牧科技职业学院 | Application of mangiferin |
| CN116617238B (en) * | 2023-05-29 | 2024-08-09 | 中国科学院水生生物研究所 | Application of Chinese herbal medicine saffron glycoside I in fish antiviral |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101461820A (en) * | 2008-12-26 | 2009-06-24 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Application of mangiferin in preparing anti-influenza virus medicament |
| CN101991026B (en) * | 2010-11-24 | 2012-02-29 | 广州海因特生物技术有限公司 | Complex immunity reinforcing agent for improving stress resistance of black carp and preparation method thereof |
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2015
- 2015-07-24 CN CN201510444307.5A patent/CN104997769A/en active Pending
- 2015-07-24 CN CN201710855423.5A patent/CN107468682B/en active Active
Non-Patent Citations (3)
| Title |
|---|
| BEN-PING ZHANG等: "Mangiferin activates Nrf2-antioxidant response element signaling without reducing the sensitivity to etoposide of human myeloid leukemia cells in vitro", 《ACTA PHARMACOLOGICA SINICA》 * |
| YI YANG等: "Up-regulation of nuclear factor E2-related factor 2 upon SVCV infection", 《FISH & SHELLFISH IMMUNOLOGY》 * |
| 廖洪利 等: "芒果苷药理研究进展", 《天津药学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114533799A (en) * | 2022-03-17 | 2022-05-27 | 湖南师范大学 | Application of callicarpa nudiflora in antiviral aspect of freshwater fishes |
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| Publication number | Publication date |
|---|---|
| CN104997769A (en) | 2015-10-28 |
| CN107468682B (en) | 2021-03-02 |
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Effective date of registration: 20210813 Address after: 430200 Huangjin 1st Road, Jinjin Industrial Park, Zhengdian street, Jiangxia District, Wuhan City, Hubei Province Patentee after: WUHAN NONGDA BIO-TECHNOLOGY Co.,Ltd. Address before: 430070 No. 1 Lion Rock street, Hongshan District, Hubei, Wuhan Patentee before: HUAZHONG AGRICULTURAL University |
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