CN110433148A - Application of the Tenovin-1 in preparation treatment and/or prevention dengue virus infection drug - Google Patents

Application of the Tenovin-1 in preparation treatment and/or prevention dengue virus infection drug Download PDF

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CN110433148A
CN110433148A CN201910839949.3A CN201910839949A CN110433148A CN 110433148 A CN110433148 A CN 110433148A CN 201910839949 A CN201910839949 A CN 201910839949A CN 110433148 A CN110433148 A CN 110433148A
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tenovin
virus
cell
protein
dengue
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姚新刚
万毅虹
刘叔文
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Southern Medical University
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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
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/17—Amides, e.g. hydroxamic acids having the group >N—C(O)—N< or >N—C(S)—N<, e.g. urea, thiourea, carmustine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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  • Organic Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention discloses a kind of application of Tenovin-1 in preparation treatment and/or prevention dengue virus infection drug, the present invention confirms that Tenovin-1 has good inhibiting effect to DENV2 by the Inhibition test to DENV, and inherently demonstrating the drug has a wide range of application background in treatment DENV2 infectious disease.The invention can provide good drug candidate for clinical treatment disease as caused by DENV2, have very good application prospect.

Description

Tenovin-1 is in preparation treatment and/or prevention dengue virus infection drug Using
Technical field
The invention belongs to pharmaceutical technology field, it is related to a kind of Tenovin-1 in preparation treatment and/or prevention dengue fever virus Application in infection medicine.
Background technique
Dengue virus (dengue virus, DENV) belongs to flaviviridae, and Flavivirus (Flaviviridae), is to work as this life It is distributed a kind of most wide arboviruse in boundary, dengue fever is mainly caused by the Aedes aegypti and aedes albopictus propagation that bite people, In to there is dengue hemorrhagic fever/dengue shock syndrome (dengue hemorrhagic fever/dengue shock Syndrome, DHF/DSS) it is even more serious.
There are four serotypes for the virus, and 2 type of serum, which is popular diseased plant dengue fever (dengue fever), to be propagated through mosquito matchmaker Acute infectious disease, mainly tropical and subtropical region prevalence.Since 1779 find dengue fever for the first time, land all over the world Continuous outburst Epidemic Situation of Dengue Fever.Estimate according to WHO, threat of the population in the whole world about 2/5 by dengue fever, there is the people of hundred million meters every year at present Infect the virus.China is in addition to there is prevalence in Taiwan, and epidemic situation is concentrated mainly on Guangdong, mostly in outburst and Introduced cases prevalence Feature, it is known that 4 kinds of serotype dengue virus be found in China, and in diffusion increase trend.
Currently, the mechanism of dengue virus infection is still indefinite, due to lacking effective cross protection between each serotype, again There are humidifications of antibody-dependant etc., and still available without safely and effectively vaccine so far, clinically there are no effective medicines Object.Although having found many bioactive molecules for being directed to dengue fever virus major protein, for various reasons (such as toxic side effect Problem), there is presently no the drugs for being directed to dengue fever virus, and therefore, the discovery for the antiviral activity molecule of dengue fever has There are important biological study meaning and practice significance, provides support to develop the active drug of dengue fever virus.
Summary of the invention
It is an object of the invention to develop a kind of drug treated and/or prevent dengue virus infection.
The purpose of the present invention is what is be achieved through the following technical solutions:
Application of the Tenovin-1 in preparation treatment and/or prevention of flavivirus viroid infection medicine.
Application of the Tenovin-1 in preparation treatment and/or prevention dengue virus infection drug.
Further, Tenovin-1 inhibits the mRNA and protein expression of key protein NS3 and E protein in dengue virus.
Further, Tenovin-1 inhibits dengue virus to generate progeny virus.
Further, dengue fever virus is dengue type 2 virus.
A kind of pharmaceutical composition treated and/or prevention of flavivirus viroid infects, described pharmaceutical composition includes conduct The Tenovin-1 of active material.
A kind of pharmaceutical composition treated and/or prevent dengue virus infection, described pharmaceutical composition include as work The Tenovin-1 of property substance.
The structure of the compound Tenovin-1 is as follows:
Compound Tenovin-1 of the present invention is the New skeleton compound delivered in 2008, which is A kind of biologically active small molecule p53 activator is a p53 activator, can prevent the p53 degradation of MDM2 mediation, mention Horizontal and inducing endogenous p53 dependence promoter the expression of high p53 and p21CIP/WAF1, shows effective in vitro Antiproliferative activity.For Tenovin-1 of the present invention in 2 type dengue virus infection drugs of preparation treatment and/or prevention In purposes belong to first public, framework types belong to completely new framework types, due to the type of compounds belong to it is non-disease-resistant Cytotoxic drug type has good effect to 2 type dengue fever virus inhibitory activity, and there is no taken the post as due to other compounds The possibility of what enlightenment has substantive distinguishing features outstanding, while obviously having for the treatment of dengue virus infection and prevention and treatment Significant progress, it is most likely that be developed to the drug of novel anti-dengue infection.
Compared with prior art, the present invention having the following advantages that and effect: the present invention passes through the Inhibition test to DENV and demonstrate,proves Real Tenovin-1 has good inhibiting effect to DENV2, inherently demonstrates the drug in treatment DENV2 infection disease There is a wide range of application background in disease.The invention can provide good candidate medicine for clinical treatment disease as caused by DENV2 Object has very good application prospect.
Detailed description of the invention
Fig. 1 is the inhibiting effect result figure of Tenovin-1 BHK-21 intracellular virus RNA metainfective to DENV2；
Fig. 2 is the inhibiting effect result of Tenovin-1 BHK-21 intracelluar toxalbumin synthesis metainfective to DENV2 Figure；
Fig. 3 is the inhibiting effect result figure that Tenovin-1 BHK-21 cell metainfective to DENV2 generates progeny virus.
Specific embodiment
Technical scheme is described further with attached drawing combined with specific embodiments below, but it is understood that this hair Bright protection scope is not limited by specific embodiment.
Embodiment 1
Tenovin-1 is 98.25% to the inhibiting rate of 2 type dengue fever virus under 10.0 μM of concentration.And Tenovin-1 exists Under the conditions of 40 μM, the survival rate to BHK-21 cell is 50%.Tenovin-1 can effectively inhibit the increasing of dengue fever virus Value, it is smaller to cytotoxicity, it can be further developed as the drug for the treatment of dengue virus infection, be with a wide range of applications. It is verified using following experiment.
1, toxicity test of the Tenovin-1 to BHK-21 cell
BHK-21 is newborn hamster kidney cell (Baby Hamster Syrian Kidney), can be by II type dengue virus (DENV2) it infects, is that dengue virus research uses more one of cell.This experiment detects Tenovin-1 to BHK-21 first The cytotoxicity of cell acts on BHK-21 cell with the Tenovin-1 of various concentration, to understand cell in different Tenovin- Survival rate under 1 concentration provides reference data to the inhibiting effect experiment of DENV2 for subsequent Tenovin-1.BHK-21 cell (newborn hamster kidney cell) is the permissive cell of DENV2.BHK-21 cell in experiment is all for this department；MTT is purchased from the green skies Biotechnology research institute；Fetal calf serum is purchased from U.S. GIBICO company；Tissue culture plate is purchased from Corning Incorporated；RPMI1640 Culture medium is purchased from U.S. GIBICO company.
Experimental procedure is as follows:
1.1 inoculation BHK-21 cells: individual cells are made into 1640 culture medium of RPMI containing 10% (V/V) fetal calf serum Suspension, with 10000, every hole cell inoculation to 96 porocyte culture plates, every hole is inoculated with 100 μ l of volume；
1.2 culture BHK-21 cells: at 37 DEG C, 5% (V/V) CO2It is cultivated 24 hours under condition of culture；
1.3 are added Tenovin-1: inhaling the culture medium abandoned in each hole, 100 μ l, 10% (V/V) tire is added to each hole 1640 culture medium of RPMI of cow's serum is diluted to the Tenovin-1 of respective concentration, and 10% (V/ of not drug containing is added in control wells V) 1640 culture medium of RPMI, the 100 μ l of fetal calf serum；
1.4 colour generations: after culture 48 hours, every hole adds 10 μ l of 5mg/ml MTT solution, at 37 DEG C, 5% (V/V) CO2Culture Under the conditions of continue culture 4 hours, DMSO is then added, until under ordinary optical microscope observation discovery Formazan all dissolve；
1.5 measurements and calculating: absorption value is measured in 570nm, the survival rate of the cell under Tenovin-1 various concentration is should Cell absorbance value is than the light absorption value of upper control wells under concentration, and multiplied by 100%, the results are shown in Table 1.
Toxicity test of the Tenovin-1 of 1 various concentration of table to BHK-21 cell
Survival rate of the Tenovin-1 under 40 μM of concentration is about 50% as the result is shown for this, in the case where being lower than the concentration, to cell Proliferation activity influences small, it was demonstrated that Tenovin-1's is highly-safe.
2, Inhibition test of the Tenovin-1 to DENV2 infection BHK-21 cell
The BHK-21 cell for having infected DENV2 is acted on the Tenovin-1 of various concentration, from cell survival rate, Virus protein is horizontal, and mRNA level in-site and daughter of virus yield four aspects obtain Tenovin-1 to the inhibition efficiency of virus. In this experiment Tenovin-1 using concentration making BHK-21 cell in the range of the concentration of 50% or more survival rate, can The analysis of experimental data is had an impact with excluding the excessive concentration of Tenovin-1.
It is the cell for cultivating virus DENV2,10TCID with BHK-2150, experimental procedure is as follows:
2.1 access BHK-21 in tissue culture plate, and cell is long to single layer after 24 hours, and cell covers the area of bottom hole about It is 80%~90%, culture medium is sucked out, PBS is washed 1 time, and 200 μ l of access viral sample, 37 DEG C adsorb 1 hour.After the completion of absorption, The virus liquid abandoned in each hole is inhaled, PBS is cleaned 1 time.It is diluted that 1640 culture medium of RPMI containing 10% (V/V) fetal calf serum is added The Tenovin-1 of prescribed concentration, in 37 DEG C of 5% (V/V) CO2It is cultivated under condition of culture.After 96 hours, cell occurs apparent thin After born of the same parents' lesion, supernatant is collected, surveys lactic dehydrogenase (LDH) content of cell release in supernatant；Or cultivate to after 48 hours, it receives Collect cell conditioned medium, does virus plaque experiment；Or cultivate to after 48 hours, the total mRNA of total protein for collecting cell passes through albumen respectively The content of the content and viral RNA of Western blot and real-time fluorescence quantitative PCR detection intracelluar toxalbumin.
2.2 survey the Dehydrogenase Content of cell release in supernatant: 96 orifice plate cell conditioned medium, 120 μ l is drawn, it is de- with lactic acid Hydrogen enzyme citotoxicity detection kit (LDH Cytotoxicity Assay Ki, Beyotime) detects cell toxicant caused by DENV The activity of the lactic dehydrogenase discharged when property, the results are shown in Table 2.
Influence of the Tenovin-1 of 2 various concentration of table to the LDH of the DENV2 BHK-21 cell release infected
Data show, Tenovin-1 has reached 80% at 5 μM to the inhibiting rate of DENV2, and 10 μM of whens have reached 98%, illustrate that Tenovin-1 inhibits making good use of for DENV2 infection cell.
3, Tenovin-1 inhibits test to the mRNA of DENV2 key protein NS3 and E protein
It include structural proteins and non-structural protein, and structural proteins E protein and NS3 gene egg in DENV2 virus It plays an important role in the white breeding amplification in DENV2 virus, therefore detects the Tenovin-1 of various concentration to NS3 and E protein MRNA suppression level, to further confirm that the inhibitory effect of Tenovin-1, the characteristic of Tenovin-1 itself can be excluded and The analysis of experimental data is had an impact.
Experimental procedure is as follows:
3.1 collect cell total rna: 1ml Trizol reagent (ambion) is added in the every hole of 6 porocyte culture plates, is placed at room temperature for After five minutes, Aspirate supernatant to 1.5ml Eppendorf manage；Every pipe adds 0.2ml chloroform, and concussion is incubated at room temperature 15 minutes, 4 DEG C 12000rpm is centrifuged 15 minutes, is drawn upper phase and is transferred to another 1.5ml Eppendorf pipe；0.5ml isopropanol, shake is added It swings, is incubated at room temperature 10 minutes, 4 DEG C of 12000rpm are centrifuged 10 minutes；It inhales and abandons supernatant, 75% (V/V) ethyl alcohol 1ml is added, washing is heavy It forms sediment, 4 DEG C of 12000rpm are centrifuged 10 minutes, are inhaled and are abandoned supernatant；It dries at room temperature and is allowed to bleach；DEPC is added and handles 20 μ l of distilled water RNA is dissolved, -80 DEG C save backup；260/280 absorbance ratio of UV spectrophotometer measuring calculates RNA concentration.
3.2 by RNA reverse transcription be cDNA: reaction system: RNA 1mg, 5 × PrimeScript RT Master Mix (TAKARA) 4 μ l, the tri-distilled water that DEPC is handled to 20 μ l of total volume.It is careful mix after 37 DEG C of 15min, 85 DEG C 5 seconds.
3.3 real-time fluorescence quantitative PCRs detect the virus envelope protein of each sample and the rna level of non-structural protein 1: anti- It answers system (10 μ l): 1 μ l of cDNA template,QPCR Master Mix (Promega) 5 μ l, positive 0.4 μ l of anti-primer, The 3.2 μ l of tri-distilled water of DEPC processing.After mixing, 95 DEG C of 10 minutes initial denaturations, 95 DEG C 15 seconds, 60 DEG C 1 minute (40 circulation).
The positive anti-primer of each target RNA of table 3
3.4 calculate: calculating RNA water of each sample relative to control group with 2- Δ Δ C т method according to the CT value of each sample It is flat, as shown in Figure 1.
2.5 μM of Tenovin-1 can significantly inhibit the level of virus structural protein E and NS3 gene in Fig. 1, At 10 μM, Tenovin-1 almost inhibits virus protein to transcribe.Show that Tenovin-1 can inhibit to close in dengue virus The mRNA level in-site of key albumen NS3 and E protein.
4, Tenovin-1 inhibits test to the protein expression of DENV2 virus key protein NS3 and E protein
It include structural proteins and non-structural protein, and structural proteins E protein and NS3 gene egg in DENV2 virus It plays an important role in the white breeding amplification in DENV2 virus, therefore detects the Tenovin-1 of various concentration to NS3 and E protein Inhibiting effect can be mutually authenticated Tenovin-1's with mRNA data to further confirm that the inhibitory effect of Tenovin-1 Inhibit the effect of DENV2 virus.
Experimental procedure is as follows:
4.1 collect total protein of cell: after BHK-21 cell drug and dengue virus (DENV2) processing, 48h mentions egg White: cold PBS washes cell, and RIPA lysate is added in the every hole of 6 orifice plates, and (inhibitors of phosphatases is added in RIPA lysate (triumphant base biology) (triumphant base biology) and protease inhibitors (triumphant base biology)) every 100 μ l of hole, scraper, which scrapes, to be collected in 1.5ml Eppendorf and manages, 4 DEG C of 12000rcf are centrifuged 15min, shift supernatant into new Eppendorf pipe.
4.2 test sample protein concentrations: protein standard substance is diluted to 0,0.0008,0.0016,0.0032 with distilled water, 0.004,0.006,0.008mg/ml gradient concentration；Protein standard substance and each 2.5 μ l of protein sample is taken to be added in 24 orifice plates, 1 × G250 working solution 1.5ml is added in every hole, makes every hole final volume 2.5ml, and room temperature is shaken 3 minutes, measures every hole with microplate reader 570nm extinction；The protein concentration in sample is calculated according to standard curve.Data are handled, the RIPA of respective volume is added in sample Lysate keeps each sample protein concentration consistent with loading buffer.100 DEG C of denaturation 5min, -20 DEG C save backup.
4.3 protein immunoblots detect the differential expression of gradient concentration Tenovin-1 treated dengue virus albumen: The configuration of 10%SDS polyacrylamide gel: being sequentially added into various reagents, configures 10% separation gel rapidly in glass spaces Perfusion, covers rapidly one layer of nothing at the time needed for glue is concentrated in outflow perfusion (the about 0.8cm of lower edge below comb) on separation gel Water-ethanol removes dehydrated alcohol layer, is blotted with filter paper after glue polymerization to be separated, continues perfusion concentration glue, plugs comb.Electrophoresis: Power on, starting carries out electrophoresis with 80V constant pressure, after dye front enters separation gel, then 120V is changed to, according to pre-dyed albumen The separation degree of Marker and the molecular weight of target protein determine electrophoresis time, general to run 75min or so.
4.4 transferring films: taking out gel, the pvdf membrane of shear gel size from glass plate, and impregnate 1min in methyl alcohol, then 5min is impregnated with transfering buffering liquid (1*tris/glycine buffer, Biorad).By gel, filter paper, pvdf membrane is all immersed in It shifts in liquid, -- cotton pad -- filter paper -- gel -- pvdf membrane -- filter paper -- cotton pad -- of installing membrane-transferring device in the following order: cathode Anode, above-mentioned article stack one by one, accurate alignment.Membrane-transferring device is placed in transferring film box, confirmation electrode is errorless, and transferring film liquid is added To entire membrane-transferring device is covered, transferring film box and top are covered with ice cube, are powered on, 100V constant pressure transferring film 70min or so, knot Shu Hou takes out pvdf membrane.
4.5 protein blockings: pvdf membrane is placed in confining liquid, and room temperature shaker impregnates 1h, to close heterogenetic antigen.Envelope After closing, TBS/TT room temperature shaker is washed film 3 times, and each 5min is then incubated for primary antibody.
The hybridization of 4.6 primary antibodies: pvdf membrane albumen is placed in etui upwardly, is added and contains Dengue viral envelope albumen (Dengue virus Envelope protein antibody, GeneTex) and non-structural protein 1 (Dengue virus NS3 glycoprotein antibody, abcam) primary antibody TBS/T configuration 5% (w/v) skim milk buffer, 4 DEG C are shaken Bed is overnight.PBST room temperature shaker washes film six times, each 5min.
The hybridization of 4.7 secondary antibodies: pvdf membrane albumen is placed in etui upwardly, be added containing secondary antibody (rabbit-anti or mouse are anti-, Proteintech 5% (w/v) skim milk buffer of TBS/T configuration), room temperature shaker 1 hour.PBST room temperature shaker is washed Film six times, each 5min.
4.8 chemiluminescences development: it is uniformly added into ECL developer solution A/B liquid (CST) in proportion, pvdf membrane is immersed in luminous In liquid, 2min is reacted.Pvdf membrane is taken out, is placed in magazine, exograph is placed in darkroom exposure, and exposure takes out development, dries glue Piece records, and saves.As shown in Figure 2.
As Tenovin-1 concentration increases in Fig. 2, the albumen of virus structural protein E protein and NS3 gene Level gradually decreases, the expression of 10 μM of complete inhibition GAP-associated protein GAPs.Show that Tenovin-1 can inhibit crucial egg in dengue virus The expression of white NS3 and E protein.
5, Tenovin-1 generates the plaque inhibition test of progeny virus to DENV2 virus
Plaque inhibition test is to detect an important indicator of virus titer and infection ability, after virus infected cell, by In the limitation of solid dielectric, the virus of release can only be from the cell that initially infects to circumferential expansion.By several proliferating cycles, just A Focal lesion cellular regions are formed, this i.e. virus plaques.After dyeing and washing, virus plaques will not catch color, To be presented as blank, the intact presentation blue of rest part cell.The processed BHK-21 cell supernatant of Tenovin-1 is taken After out, the virus plaque experiment of supernatant, after capable of further confirming Tenovin-1 processing in this way, the disease in cell conditioned medium are carried out The level of malicious content, titre and infection ability.More accurately judge the anti-DENV2 progeny virus effect of Tenovin-1.
Experimental procedure is as follows:
5.1 access C6/36 cell in 24 orifice plates, and to single layer, (cell covers the area of bottom hole about to cell length after 24 hours 90%~100%) culture medium abandoned in each hole, is inhaled, PBS is washed 1 time, the access diluted 200 μ l of viral sample of PBS, 37 DEG C of suctions Attached 1 hour.
The supernatant in each hole is inhaled after the completion of 5.2 absorption and is abandoned, PBS washes away unadsorbed virus.It is added fine containing 1.2% methyl 1640 culture medium of RPMI for tieing up plain 2% (V/V) fetal calf serum, in 37 DEG C, 5% (V/V) CO2Incubator in cultivate 96~120 Hour.
5.3 inhale abandoning methylcellulose covering culture medium after plaque test is fixed with 4% paraformaldehyde (doctor's moral biology), Again with 1% (w/v) violet staining, crystal violet is washed away with flowing water after 2 hours, 24 orifice plates are scanned after drying, is sentenced according to plaque number Disconnected Mol-5 BHK-21 cell metainfective to DENV2 generates the inhibiting effect of progeny virus.As shown in Figure 3.
As Tenovin-1 concentration increases in Fig. 3, daughter of virus levels of replication is gradually decreased, at 10 μM daughter of virus without Method release.Show that Tenovin-1 can inhibit dengue virus filial generation levels of replication.
The foregoing is merely a specific embodiment of the invention, the embodiment being not all of, ordinary skill people Any equivalent transformation that member takes technical solution of the present invention by reading description of the invention, is right of the invention It is required that being covered.
SEQUENCE LISTING
<110>Nanfang Medical Univ
<120>application of the Tenovin-1 in preparation treatment and/or prevention dengue virus infection drug
<130> CP11901472C
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Claims (7)

  1. Application of the 1.Tenovin-1 in preparation treatment and/or prevention of flavivirus viroid infection medicine.
  2. Application of the 2.Tenovin-1 in preparation treatment and/or prevention dengue virus infection drug.
  3. 3. application according to claim 2, which is characterized in that the Tenovin-1 inhibits key protein in dengue virus The mRNA and protein expression of NS3 and E protein.
  4. 4. application according to claim 2, which is characterized in that the Tenovin-1 inhibits dengue virus to generate filial generation disease Poison.
  5. 5. according to any application of claim 2-4, which is characterized in that the dengue fever virus is dengue type 2 virus.
  6. 6. the pharmaceutical composition of a kind for the treatment of and/or the infection of prevention of flavivirus viroid, which is characterized in that described pharmaceutical composition Including the Tenovin-1 as active material.
  7. 7. the pharmaceutical composition of a kind for the treatment of and/or prevention dengue virus infection, which is characterized in that described pharmaceutical composition packet Include the Tenovin-1 as active material.
CN201910839949.3A 2019-09-06 2019-09-06 Application of the Tenovin-1 in preparation treatment and/or prevention dengue virus infection drug Pending CN110433148A (en)

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Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BRENT A.等: "Sirtuin inhibitors are broadly antiviral against arboviruses.", 《MBIO》 *
RAUSCH, KEIKO 等: "Screening Bioactives Reveals Nanchangmycin as a Broad Spectrum Antiviral Active against Zika Virus.", 《CELL REPORTS》 *

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Application publication date: 20191112