CN108478562A - The application of Mycophenolic Acid and its derivative mycophenolate mofetil in preparing wide spectrum anti-coronavirus drug - Google Patents
The application of Mycophenolic Acid and its derivative mycophenolate mofetil in preparing wide spectrum anti-coronavirus drug Download PDFInfo
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Abstract
The invention discloses the application of Mycophenolic Acid and its derivative mycophenolate mofetil in preparing wide spectrum anti-coronavirus drug.The present invention selects the Mycophenolic Acid of non-toxic concentrations and its derivative mycophenolate mofetil to carry out wide spectrum anti-coronavirus research, it was found that the two can effectively inhibit the duplication of β crowds of coronavirus HCoV OC43, A59 and α crowds of MERS CoV, MHV coronavirus HCoV NL63 in vitro, and dosage effect correlation is presented, Mycophenolic Acid for the HCoV OC43 of MOI=0.01 dosage, HCoV NL63, MERS CoV and MHV tetra- kinds of coronavirus of A59 EC50Respectively 1.65uM, 0.18uM, 1.81uM and 0.17uM；Mycophenolate mofetil for the HCoV OC43 of MOI=0.01 dosage, HCoV NL63, MERS CoV and MHV tetra- kinds of coronavirus of A59 EC50Respectively 1.45uM, 0.23uM, 1.98uM and 0.27uM；It shows that Mycophenolic Acid and its derivative mycophenolate mofetil have wide spectrum anti-coronavirus activity, lays a good foundation for further exploitation wide spectrum anti-coronavirus drug, there is important Development volue and be widely applied foreground.
Description
The invention belongs to be based on biomedicine technical field more particularly to Mycophenolic Acid and its derivative for technical field
For new opplication of the mycophenolate in preparing wide spectrum anti-coronavirus drug, and in particular to Mycophenolic Acid and its derivative replace wheat
Phenolic ester is examined in anti-Middle East breathing syndrome coronavirus, human corona virus OC43, human corona virus NL63, mouse hepatitis virus
Application in A59 drugs.
Background technology coronavirus (Coronavirus, CoV) is tunicary single strand plus RNA virus, can cause to move
The infection of the object and respiratory tract of people, alimentary canal and nervous system is a kind of important zoonosis virus.According to coronavirus
Evolution feature, the 9th report of International Commission on Virus Classification be classified as tetra- groups of α, β, γ and δ.Wherein, α with β groups
Coronavirus mainly infects mammal, γ and the δ groups of main infected poultries of coronavirus.
Coronavirus can infect people and many animals, and it is comprehensive from common cold to serious acute respiration that people can be caused to suffer from
The diseases such as simulator sickness.The coronavirus that can infect people being currently known has six kinds, including upper respiratory tract infection symptoms is caused commonly to be felt
Human corona virus 229E (HCoV-229E), NL63 (HCoV-NL63), HKU1 (HCoV-HKU1), the OC43 (HCoV-OC43) emitted
And atypical pneumonia coronavirus (the Severe acute of severe breathing problem can be led to
Respiratory syndrome coronavirus, SARS-CoV) and Middle East respiration syndrome coronavirus (Middle
East respiratory syndrome coronavirus, MERS-CoV).Wherein, two kinds of highly pathogenic coronavirus
SARS-CoV and MERS-CoV belongs to β groups of coronavirus, this group of viruses further include HCoV-HKU1, HCoV-OC43 and conduct
Cause of disease mouse hepatitis virus (the Murine of people's hepatitis and fatty degeneration of liver and the subacute important animal model of cental system pathology
Hepatitis virus, MHV) etc.；HCoV-229E, HCoV-NL63 that the disease symptoms of initiation are lighter belong to α groups of coronal diseases
Poison.It is worth noting that although coronavirus is one of the common causative for causing acute human respiratory tract infection, mesh
Before there is no the specific drug of approval to treat infection caused by coronavirus, even if being infected for by SARS-CoV and MERS-CoV
Caused serious acute respiratory patient, clinic is also mainly based on symptomatic treatment reduction complication for patients.Therefore, there is an urgent need for develop
To treat, coronavirus is caused to be infected effective drug.
Monoclonal antibody, polypeptide and micromolecular compound are typically the hot spot of antiviral drugs research, by having been criticized to FDA
Accurate drug reservoir carries out high flux screening, inquires into the new application for having drug, it has also become a kind of important channel of medicament research and development.By
Have the function of, about data such as pharmacological effect test, target spot and clinical safeties, to be conducive to further in such as drug candidate
Toxicological evaluation, Pharmacokinetic Evaluation and preparation research and development etc., R＆D risk can be greatly reduced, shorten research and development and the time and grind
Cost is sent out, is had broad application prospects.Antiviral drugs for MERS-CoV is studies have shown that by Interferon Alpha-2b
(interferon alfa-2b) and Ribavirin (ribavirin) drug combination can inhibit in cultured cell in vitro
The duplication of MERS-CoV, and lung virus carrying capacity can be reduced and mitigate the immune response of host, and this conjoint therapy can carry
14 days survival rates of high patient.However, the conjoint therapy may be only effective to the patient of certain early diagnosis, to severe infections
And the late diagnosis patient containing multiple complications is invalid.Recently the study found that treatment Ebola virus infection nucleic acid
There is broad anti-viral activity like object pro-drug GS-5734, also can inhibit MERS-CoV in cell culture model in vitro
Duplication, provide a new direction for the treatment of MERS.In addition, MERS-CoV specific polypeptides inhibitor, chloroquine, chlorine third
The broad-spectrum medicinals such as piperazine, mycophenolic acid and Nitazoxanide also can suppressing virus replication, but clinically whether effectively, need it is further true
Recognize.
Mycophenolic Acid (mycophenolic acid, MPA), also referred to as mycophenolic acid, be generated by Penicillium bacterial strain it is anti-true
Bacterium, antibiotic that is antitumor and having immunosuppressive action；2- ethyl ester derivatives --- the mycophenolate mofetil of Mycophenolic Acid
(mycophenolate mofetil, MMF), also referred to as mycophenolate are that one kind of Switzerland's Roche (Roche) company research and development is novel
Immunosuppressor lists in the U.S. for the first time in nineteen ninety-five, and trade name learn (Cellcept) forms tool in vivo after esterification
The metabolite Mycophenolic Acid for having immunosuppressive activity, by the crucial speed limit for inhibiting purine nucleotides de novo synthesis
Enzyme --- inosine phosphate dehydrogenase reduces the synthesis of guanylic acid, thus energy selective depression and repulsion are anti-
The proliferation and function for answering related T, bone-marrow-derived lymphocyte, are mainly used for the prevention of organ rejection after kidney, heart transplant.
Studies have shown that Mycophenolic Acid have broad anti-viral activity, can effectively inhibit in vitro include west nile virus,
The infection of yellow fever virus, chikungunya fever virus, Hepatitis C Virus and MERS-CoV.MERS-CoV antiviral drugs high passes
It measures screening study to find, medium effective concentration (the Concentration for 50%of maximal of Mycophenolic Acid
Effect, EC50) it is 0.24uM, half cytotoxic concentration (Concentration cytotoxicity 50%, CC50) be
170uM selects index (Slectivity index, SI, SI=CC50/EC50, SI ＞ 1 are effective) be up to 708 (Chan JF,
Chan KH, Kao RY, et al.Broad-spectrum antivirals for the emerging Middle East
Respiratory syndrome coronavirus.J Infect, 2013,67,606-616.).Compare MERS-CoV confrontation
The sensibility of virus drugs (Mycophenolic Acid and Ribavirin) and difference IFN products the study found that IFN-β is shown pair
The strongest external inhibitory activity of MERS-CoV, EC50For 1.37U/ml；Ribavirin not table under the dosage of Current treatment protocols
Reveal HIV suppression activity；In contrast, Mycophenolic Acid shows stronger inhibiting effect, EC50For 2.87 μm of ol/L (Hart
BJ, Dyall J, Postnikova E, et al.Interferon- β and mycophenolic acid are potent
inhibitors of Middle East respiratory syndrome coronavirus in cell-based
Assays.J Gen Virol, 2014,95,571-577).The studies above prompt, Mycophenolic Acid and IFN-β, which are applied alone or combine, to be made
Therapeutic intervention that may be to MERS-CoV the infected or as high exposed population group is used to play preferably effect.But at present not yet
See the research of Mycophenolic Acid and mycophenolate mofetil as the drug of wide spectrum anti-coronavirus.
Based on above-mentioned analysis, the present invention is with β groups of coronavirus (HCoV-OC43, MERS-CoV and MHV-A59) and α groups
Coronavirus (HCoV-NL63) is used as model virus, by extracorporeal antivirus effect effect disquisition, inquires into Mycophenolic Acid and its derivative
The possibility of application of the mycophenolate mofetil in preparing wide spectrum anti-coronavirus drug finds Mycophenolic Acid and its derivative
HCoV-OC43, MERS-CoV, MHV-A59 and α groups of coronal diseases of β groups of coronavirus can be significantly inhibited in vitro for mycophenolate
The duplication of malicious HCoV-NL63 is of great significance for the prevention and control and treatment of coronavirus infection.
The present invention provides a kind of Mycophenolic Acids and its derivative mycophenolate mofetil to prepare the anti-hat of wide spectrum for invention content
New opplication in shape virus drugs, it was confirmed that Mycophenolic Acid and its derivative mycophenolate mofetil can have in cell model in vitro
Effect inhibits the nucleic acid of HCoV-OC43, MERS-CoV, MHV-A59 and α crowds of coronavirus HCoV-NL63 of β groups of coronavirus multiple
System, the drug for further exploitation wide spectrum anti-coronavirus are laid a good foundation.
Description of the drawings
Fig. 1 is Mycophenolic Acid and its chemical molecular structural schematic diagram of derivative mycophenolate mofetil.Figure 1A is that wheat examines phenol
The chemical molecular structural schematic diagram of acid, Figure 1B are the chemical molecular structural schematic diagram of mycophenolate mofetil.
Fig. 2 is four kinds of coronavirus antiviral effects of various concentration Mycophenolic Acid and its derivative mycophenolate mofetil pair.
Fig. 2A is various concentration Mycophenolic Acid to the inhibition to HCoV-NOC43 virus replications.Abscissa is that wheat examines phenol
The drug concentration of acid, ordinate is inhibiting rate, and the curve of wherein box dark color point fitting indicates the virus replication to HCoV-OC43
Efficiency, the lighter curve of circular solids point fitting is inhibited to indicate the inhibition efficiency to BHK-21 cell activity.The results show that wheat is examined
Phenolic acid can effectively inhibit the virus replication of HCoV-OC43 in BHK-21 cells, make the HCoV-OC43 diseases of MOI=0.01 dosage
The drug medium effective concentration EC that malicious duplicating efficiency is inhibited by 50%50For 1.65uM.
Fig. 2 B are various concentration Mycophenolic Acid to the inhibition to HCoV-NL63 virus replications.Abscissa is that wheat examines phenol
The drug concentration of acid, ordinate is inhibiting rate, and the curve of wherein box dark color point fitting indicates the virus replication to HCoV-NL63
Efficiency, the lighter curve of circular solids point fitting is inhibited to indicate the inhibition efficiency to LLC-MK2 cell activity.The results show that wheat
The virus replication of HCoV-NL63 can effectively be inhibited in LLC-MK2 cells by examining phenolic acid, make the HCoV-NL63 of MOI=0.01 dosage
The drug medium effective concentration EC that viral replication efficiency is inhibited by 50%50For 0.18uM.
Fig. 2 C are inhibition of the various concentration Mycophenolic Acid to MERS-CoV virus replications.Abscissa is Mycophenolic Acid
Drug concentration, ordinate are inhibiting rate, and the curve of wherein box dark color point fitting indicates to inhibit the virus replication of MERS-CoV
The lighter curve of efficiency, the fitting of circular solids point indicates the inhibition efficiency to Vero-E6 cell activity.The results show that wheat examines phenol
Acid can effectively inhibit the virus replication of MERS-CoV in Vero-E6 cells, keep the MERS-CoV viruses of MOI=0.01 dosage multiple
The drug medium effective concentration EC that efficiency processed is inhibited by 50%50For 1.81uM.
Fig. 2 D are inhibition of the various concentration Mycophenolic Acid to MHV-A59 virus replications.Abscissa is Mycophenolic Acid
Drug concentration, ordinate are inhibiting rate, and the curve of wherein box dark color point fitting indicates to inhibit effect to the virus replication of MHV-A59
The lighter curve of rate, the fitting of circular solids point indicates the inhibition efficiency to DBT cell activity.The results show that Mycophenolic Acid is in DBT
The virus replication that MHV-A59 can effectively be inhibited in cell, make the MHV-A59 viral replication efficiencies of MOI=0.01 dosage by
The 50% drug medium effective concentration EC inhibited50For 0.17uM.
Fig. 2 E are various concentration mycophenolate mofetil to the inhibition to HCoV-NOC43 virus replications.Abscissa be
For the drug concentration of mycophenolate, ordinate is inhibiting rate, and the curve of wherein box dark color point fitting is indicated to HCoV-OC43's
Virus replication inhibits efficiency, the lighter curve of circular solids point fitting to indicate the inhibition efficiency to BHK-21 cell activity.As a result
It has been shown that, mycophenolate mofetil can effectively inhibit the virus replication of HCoV-OC43 in BHK-21 cells, make MOI=0.01 dosage
HCoV-OC43 viral replication efficiencies by 50% inhibit drug medium effective concentration EC50For 1.45uM.
Fig. 2 F are various concentration mycophenolate mofetil to the inhibition to HCoV-NL63 virus replications.Abscissa be
For the drug concentration of mycophenolate, ordinate is inhibiting rate, and the curve of wherein box dark color point fitting is indicated to HCoV-NL63's
Virus replication inhibits efficiency, the lighter curve of circular solids point fitting to indicate the inhibition efficiency to LLC-MK2 cell activity.As a result
It has been shown that, mycophenolate mofetil can effectively inhibit the virus replication of HCoV-NL63 in LLC-MK2 cells, make MOI=0.01 dosage
HCoV-NL63 viral replication efficiencies by 50% inhibit drug medium effective concentration EC50For 0.23uM.
Fig. 2 G are inhibition of the various concentration mycophenolate mofetil to MERS-CoV virus replications.Abscissa is to replace wheat
The drug concentration of phenolic ester is examined, ordinate is inhibiting rate, and the curve of wherein box dark color point fitting indicates the virus to MERS-CoV
The lighter curve of inhibition of DNA replication efficiency, the fitting of circular solids point indicates the inhibition efficiency to Vero-E6 cell activity.As a result it shows
Show, mycophenolate mofetil can effectively inhibit the virus replication of MERS-CoV in Vero-E6 cells, make MOI=0.01 dosage
The drug medium effective concentration EC that MERS-CoV viral replication efficiencies are inhibited by 50%50For 1.98uM.
Fig. 2 H are inhibition of the various concentration mycophenolate mofetil to MHV-A59 virus replications.Abscissa is to replace wheat
The drug concentration of phenolic ester is examined, ordinate is inhibiting rate, and the curve of wherein box dark color point fitting indicates multiple to the virus of MHV-A59
System inhibits efficiency, the lighter curve of circular solids point fitting to indicate the inhibition efficiency to DBT cell activity.The results show that replacing
Mycophenolate can effectively inhibit the virus replication of MHV-A59 in DBT cells, keep the MHV-A59 viruses of MOI=0.01 dosage multiple
The drug medium effective concentration EC that efficiency processed is inhibited by 50%50For 0.27uM.
Specific implementation mode：
With reference to specific embodiment, the present invention is described in further detail, it is described be explanation of the invention and
It is not to limit.Test method without specific conditions in following embodiment, usually according to normal condition such as《Molecular Cloning: A Laboratory
Guide》Condition described in reference books commonly used in the art such as (third edition, Science Presses, 2005), or press reagent manufacturer
Proposed condition carries out.
Embodiment 1：The cytotoxicity of Mycophenolic Acid and mycophenolate mofetil detects
The sensitivity of four kinds of coronavirus HCoV-OC43, HCoV-NL63, MERS-CoV and MHVA59 involved in the present invention
Cell line is BHK-21, LLC-MK2, Vero-E6 and DBT respectively, in order to test the safety of Mycophenolic Acid and mycophenolate mofetil
Drug level, this research have detected toxicity of the Mycophenolic Acid with mycophenolate mofetil to these four cell lines using mtt assay.Detection
Principle is：The bluish violet that succinate dehydrogenase in living cells mitochondria can make exogenous MTT be reduced to water-insoluble crystallizes first
Za is simultaneously deposited in cell, and dead cell is without this function.Dimethyl sulfoxide (DMSO) (Dimethylsulfoxide, DMSO) can dissolve carefully
Formazan in born of the same parents measures its absorbance value with microplate reader at respective wavelength, and within the scope of certain cell number, MTT crystallizes to be formed
Amount it is directly proportional to cell number.Judge living cells quantity according to the absorbance value (OD values) measured, OD values are bigger, cell activity
It is stronger, then indicate that drug toxicity is smaller when surveying drug toxicity.
Concrete operations are as follows：By tetra- kinds of cells of BHK-21, LLC-MK2, Vero-E6 and DBT respectively according to 1 × 104Cell/
Hole is inoculated in 96 well culture plates, in blocks with the DMEM cultures 16 hours to 80% containing 10% fetal calf serum；It inhales later and abandons culture solution,
Change the DMEM culture mediums containing 2% fetal calf serum into；Mycophenolic Acid is purchased from the sincere biotechnology of Beijing standing grain show with mycophenolate mofetil to be had
Limit company, Mycophenolic Acid come from Selleck.cn, and article No. S2487-50MG, purity ＞ 98%, mycophenolate mofetil is purchased from
Sigma-aldrich companies, article No. Y0000489, purity ＞ 98%；After using DMSO dissolvings respectively, further use PBS dilute
It releases, Mycophenolic Acid and mycophenolate mofetil is pressed into final concentration of 0.5uM, 2uM, 5uM, cell, every group of acute drug is added in 10uM
Do three groups it is parallel, while blank control group and cell controls group are set.37 DEG C are positioned over, 5%CO2It is small to continue culture 72 in incubator
When, the 20 μ L of MTT solution with the PBS 5g/L prepared are added per hole, continue to cultivate 4h.Supernatant is carefully discarded later, per hole
Sediment, and the mixing 30min in 96 hole plate oscillators are dissolved with 100 μ L of isopropanol, is finally existed using multi-function microplate reader
Its absorbance value is measured under 570nm wavelength.It is calculated according to formula：Cytoactive inhibiting rate (%)=(medicine group-blank control
Group)/(cell controls group-blank group) × 100%.Using Mycophenolic Acid and the drug concentration of mycophenolate mofetil as abscissa, with
Cell proliferation inhibition rate is ordinate, calculates average value by 7 softwares of Graphpad Prism and standard deviation matched curve is made
Drug concentration is converted to the cytotoxicity CC of calculating Mycophenolic Acid and mycophenolate mofetil after logarithm by figure50.As a result such as Fig. 2
Shown in the lighter curve of circular solids point fitting, CC of the Mycophenolic Acid in BHK-21, LLC-MK2, Vero-E6 and DBT cell50
Respectively 4.55uM, 5.04uM, 4.15uM and 5.37uM；Mycophenolate mofetil is in BHK-21, LLC-MK2, Vero-E6 and DBT
CC in cell50Respectively 3.51uM, 4.30uM, 3.84uM and 4.45uM.
Embodiment 2：Mycophenolic Acid and the external anti-coronavirus active effect of mycophenolate mofetil detect
2.1 virus infection and drug effect
By BHK21, CCL-MK2, Vero-E6 and DBT cell respectively according to 1 × 104Cells/well is inoculated in 96 well culture plates
In, it is in blocks using the DMEM cultures 16 hours to 80% containing 10% fetal calf serum；It inhales later and abandons cell culture fluid, change into containing 2%
The DMEM culture mediums of fetal calf serum；By final concentration it is respectively 0.5uM, 2uM, 5uM, 10uM by Mycophenolic Acid and mycophenolate mofetil
Dosage be added in corresponding cell hole, while the cell control well for being accordingly not added with drug and only plus viral virus control is set
Hole；Be added drug after in 1 hour, respectively in corresponding sensitive cells hole with the volume of every hole 10ul be added HCoV-OC43,
HCoV-NL63, MERS-CoV and MHV-A59, make viral infection multiplicity be 0.01 (Multiplicity of infection,
MOI=0.01), 37 DEG C are positioned over, 5%CO2Supernatant is collected after being cultivated 72 hours in incubator.
2.2 fluorescence quantitative RT-RCRs detect inhibition of the drug to HCoV-OC43, HCoV-NL63 and MERS-CoV
It is accordingly sick using the detection of absolute fluorescence quantitative approach RT-PCR method HCoV-OC43, HCoV-NL63 and MERS-CoV
Malicious target gene transcriptional level so as to reflect virus levels of replication.According to Qiagen Viral RNA Mini Kit specifications
Viral RNA is extracted, RT-PCR detections are carried out after extracting viral RNA.Wherein, the primer probe sequence for detecting each virus is as follows：
The primer probe sequence for detecting HCoV-OC43 is as follows：
Upstream primer sequence (q-OC43-F) is：5 '-GCTCAGGAAGGTCTGCTCC-3 ',
Downstream primer sequence (q-OC43-R) is：5 '-TCCTGCACTAGAGGCTCTGC-3 ',
Probe sequence (q-OC43-probe) is：5’-TTCCAGATCTACTTCGCGCACATCC-3’.
The primer probe sequence for detecting HCoV-NL63 is as follows：
Upstream primer sequence (q-NL63-F) is：5 '-AGGACCTTAAATTCAGACAACGTTCT-3 ',
Downstream primer sequence (q-NL63-R) is：5 '-GATTACGTTTGCGATTACCAAGACT-3 ',
Probe sequence (q-NL63-probe) is：5’-AACAGTTTTAGCACCTTCCTTAGCAACCCAAACA-3’.
The primer probe sequence for detecting MERS-CoV is as follows：
Upstream primer sequence (q-MERS-F) is：5 '-GGCACTGAGGACCCACGTT-3 ',
Downstream primer sequence (q-MERS-R) is：5 '-TTGCGACATACCCATAAAAGCA-3 ',
Probe sequence (q-MERS-probe) is：5’-CCCCAAATTGCTGAGCTTGCTCCTACA-3’.
Reaction system is：12.5μL 2×One Step SYBR RT-PCR Buffer III、0.5μL Takara Ex
Taq HS, 0.5 μ L PrimeScript RT Enzyme Mix II, 1.5 μ L sense primers, 1.5 μ L downstream primers, 2 μ L RNA
Template is used in combination aseptic double-distilled water to mend to 25 μ L.Response parameter is：42 DEG C of 5min, 95 DEG C of 10s, mono- cycle；95℃ 5s、60
DEG C 30s recycles 40 times, fluorescence signal is acquired after extension.Each sample does 3 repetitions, finally counts sample CT values, will be surveyed
Sample CT values substitute into standard curve after calculate viral copy number in sample.It is calculated according to formula：Virus replication inhibiting rate
(%)=(virus control group-drug shines group)/virus control group × 100%.
2.3 plaques reduce the inhibition for inhibiting testing inspection Mycophenolic Acid with mycophenolate mofetil to MHV-A59 viruses
The inhibition of DNA replication effect that experiment measures drug to MHV-A59 viruses is reduced using plaque.By DBT according to 1 × 105Carefully
The amount in born of the same parents/hole is inoculated in 6 well culture plates, using the DMEM medium cultures 16 hours containing 10% fetal calf serum to 80% one-tenth
Piece；It inhales later and abandons cell culture fluid, change the DMEM culture mediums containing 2% fetal calf serum into；By Mycophenolic Acid and mycophenolate mofetil
It is respectively 0.5uM, 2uM, 5uM according to final concentration, the dosage of 10uM is added in corresponding cell hole, while setting is accordingly not added with drug
Cell control well and only plus virus virus control wells；It is added after drug in 1 hour, the virus that 10ul has diluted is added per hole
MHV-A59 storing liquids make viral MOI=0.01, are positioned over 37 DEG C, 5%CO2Supernatant is collected after being cultivated 72 hours in incubator,
Experiment is reduced using plaque and measures drug to the inhibition of DNA replication effect of virus, and the supernatant of collection is infected according to different dilutions
DBT cells are inhaled after 72h and abandon supernatant, and 4% paraformaldehyde uses violet staining, counting orifice to be calculated according to formula after fixing：
Virus replication inhibiting rate (%)=(virus control group-drug shines group)/virus control group × 100%.
2.4 matched curves and calculating EC50
Passed through using virus replication inhibiting rate as ordinate with for a concentration of abscissa of mycophenolate with Mycophenolic Acid
7 softwares of Graphpad Prism calculate the average value for inhibiting efficiency and standard deviation matched curve mapping, and drug concentration is converted into
Mycophenolic Acid and its EC of derivative mycophenolate mofetil are calculated after logarithm50, as a result such as the curve institute of Fig. 2 boxes dark color point fitting
Show.Mycophenolic Acid distinguishes the inhibition of tetra- kinds of coronavirus of HCoV-OC43, HCoV-NL63, MERS-CoV and MHV-A59
See Fig. 2A, 2B, 2C and 2D；Mycophenolate mofetil is coronal to tetra- kinds of HCoV-OC43, HCoV-NL63, MERS-CoV and MHV-A59
The inhibition of virus is shown in Fig. 2 E, 2F, 2G and 2H respectively.As shown in Figure 2 A, Mycophenolic Acid can effectively press down in BHK-21 cells
The duplication of HCoV-OC43 processed, the drug medium effective concentration EC for making the HCoV-OC43 duplicating efficiencies 50% of MOI=0.01 inhibit50
For 1.65uM；As shown in Figure 2 B, Mycophenolic Acid can effectively inhibit the duplication of HCoV-NL63 in LLC-MK2 cells, make
The drug medium effective concentration EC that the HCoV-NL63 duplicating efficiencies 50% of 0.01MOI inhibit50For 0.18uM；As shown in Figure 2 C, wheat
The duplication of MERS-CoV can effectively be inhibited in Vero-E6 cells by examining phenolic acid, make the MERS-CoV duplicating efficiencies of MOI=0.01
The 50% drug medium effective concentration EC inhibited50For 1.81uM；As shown in Figure 2 D, Mycophenolic Acid can effectively press down in DBT cells
The duplication of MHV-A59 processed makes the MHV-A59 of MOI=0.01 replicate the 50% drug medium effective concentration EC inhibited50For
0.17uM.As shown in Figure 2 E, mycophenolate mofetil can effectively inhibit the duplication of HCoV-OC43 in BHK-21 cells, make MOI=
The drug medium effective concentration EC that 0.01 HCoV-OC43 duplicating efficiencies 50% inhibit50For 1.45uM；As shown in Figure 2 F, it replaces
Mycophenolate can effectively inhibit the duplication of HCoV-NL63 in LLC-MK2 cells, make the HCoV-NL63 duplicating efficiencies of 0.01MOI
The 50% drug medium effective concentration EC inhibited50For 0.23uM；As shown in Figure 2 G, mycophenolate mofetil is in Vero-E6 cells
The duplication that MERS-CoV can effectively be inhibited keeps the drug half that the MERS-CoV duplicating efficiencies 50% of MOI=0.01 inhibit effective
Concentration EC50For 1.98uM；As illustrated in figure 2h, mycophenolate mofetil can effectively inhibit the duplication of MHV-A59 in DBT cells, make
The MHV-A59 of MOI=0.01 replicates the 50% drug medium effective concentration EC inhibited50For 0.27uM.
Claims (2)
1. the invention discloses a kind of Mycophenolic Acids and its derivative mycophenolate mofetil to prepare wide spectrum anti-coronavirus drug
In application, which is characterized in that the coronavirus includes：Middle East breathing syndrome coronavirus (MERS-CoV), people's hat
Shape virus O C43 (HCoV-OC43), human corona virus NL63 (HCoV-NL63), a59 virus (MHV-A59).
2. a kind of Mycophenolic Acid described in accordance with the claim 1 and its derivative mycophenolate mofetil are in wide spectrum anti-coronavirus medicine
Application in object.
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| CN201810418494.3A CN108478562B (en) | 2018-05-04 | 2018-05-04 | Application of mycophenolic acid and mycophenolate mofetil derivative thereof in preparation of broad-spectrum anti-coronavirus medicines |
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| CN201810418494.3A CN108478562B (en) | 2018-05-04 | 2018-05-04 | Application of mycophenolic acid and mycophenolate mofetil derivative thereof in preparation of broad-spectrum anti-coronavirus medicines |
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| CN113491699A (en) * | 2020-04-01 | 2021-10-12 | 中国科学院上海药物研究所 | Application of mycophenolic acid or combination preparation containing mycophenolic acid in resisting coronavirus |
| CN114053266A (en) * | 2021-12-29 | 2022-02-18 | 佛山科学技术学院 | Application of mycophenolic acid in preparation of medicine for preventing and treating Porcine Reproductive and Respiratory Syndrome (PRRSV) |
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| CN115252610A (en) * | 2020-02-26 | 2022-11-01 | 上海科技大学 | Application of proton pump inhibitor in resisting coronavirus |
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| CN112315962A (en) * | 2020-11-17 | 2021-02-05 | 北京化工大学 | Application of mycophenolic acid in treating and preventing coxsackie virus infection |
| CN112315962B (en) * | 2020-11-17 | 2022-03-29 | 北京化工大学 | Application of mycophenolic acid in treating and preventing coxsackie virus infection |
| CN114053266A (en) * | 2021-12-29 | 2022-02-18 | 佛山科学技术学院 | Application of mycophenolic acid in preparation of medicine for preventing and treating Porcine Reproductive and Respiratory Syndrome (PRRSV) |
| WO2023150067A1 (en) * | 2022-02-03 | 2023-08-10 | Steven Baranowitz | Pharmaceutical compositions and methods for treating covid |
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