CN117379431A - Application of pyrimidine derivative in preparation of medicines for resisting porcine reproductive and respiratory syndrome virus - Google Patents
Application of pyrimidine derivative in preparation of medicines for resisting porcine reproductive and respiratory syndrome virus Download PDFInfo
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- KEYDJKSQFDUAGF-YIRKRNQHSA-N prostaglandin D2 ethanolamide Chemical compound CCCCC[C@H](O)\C=C\[C@@H]1[C@@H](C\C=C/CCCC(=O)NCCO)[C@@H](O)CC1=O KEYDJKSQFDUAGF-YIRKRNQHSA-N 0.000 description 1
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Classifications
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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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
-
- 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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4418—Non condensed pyridines; Hydrogenated derivatives thereof having a carbocyclic group directly attached to the heterocyclic ring, e.g. cyproheptadine
-
- 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
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- Health & Medical Sciences (AREA)
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- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Virology (AREA)
- Molecular Biology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides an application of pyrimidine derivatives in preparing medicines for resisting porcine reproductive and respiratory syndrome viruses, and belongs to the field of chemical medicines. The pyrimidine derivative has a structure shown in a formula I. Experimental results show that the compound has remarkable inhibition and blocking effects on the-1-position ribosome frameshift (-1 Ribosomal Frameshifting) process of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), can effectively inhibit the replication of PRRSV, inhibit the proliferation of PRRSV, and the 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine can be used for preparing anti-PRRSV medicaments and medicaments for treating blue pig ear diseases caused by PRRSV, and has wide application prospects.
Description
Technical Field
The invention belongs to the field of chemical medicines, and particularly relates to application of a pyrimidine derivative in preparation of a medicine for resisting porcine reproductive and respiratory syndrome virus.
Background
Porcine Reproductive and Respiratory Syndrome (PRRS), also known as blue pig ear disease, is an acute infectious disease caused by porcine reproductive and respiratory syndrome virus (porcine reproductive and respiratory syndrome virus, PRRSV), which affects mainly the respiratory system of pigs (characterized by interstitial pneumonia), causing abortion in sows, increased incidence of stillbirth, fetal collapse and weakness of newborn piglets and slow growth of piglets. In addition to resulting in a reduced number of pigs, significant manpower and resources are also taken up in order to control and contain such infectious diseases, and PRRSV therefore causes significant losses to the pig industry.
PRRSV belongs to the genus arterivirus of the family arterividae, the virus diameter is about 50-60nm, the genome is a single-stranded sense strand RNA, the length is 15kb, and it contains 11 Open Reading Frames (ORFs). Wherein the two open reading frames upstream are ORF1a and ORF1b, respectively, which occupy two thirds of the genome and contain at least 1ribosomal frameshift site. ORF1a and ORF1b encode two large nonstructural polyproteins: pp1a, pp1ab, wherein expression of pp1ab depends on the-1-position ribosomal frameshift (-1 Ribosomal Frameshifting) element of the ORF1a/ORF1b overlap region. After synthesis of pp1a and pp1ab replicase polyproteins from genomic RNA, they were processed into at least 14 nonstructural proteins (nsps) by 4 proteases encoded by ORF1a located in nsp1α, nsp1β, nsp2 and nsp 4.
The "-1Ribosomal Frameshifting" phenomenon in ribosome elongation is very common in RNA viruses, and is the phenomenon in IBV, HIV-1 and SARS viruses, and the specific process is as follows: (1) The pseudo-junction structure of mRNA forces ribosome to arrest in the extension phase, and the anticodon loops of the A-site aminoacyltRNA and the P-site peptidyl tRNA are just combined with the sliding sequence of mRNA; (2) the sliding sequence causes a shift in tRNAs at position-1; (3) Downstream mRNA pseudojunctions are opened and the ribosome continues to move forward, but the reading frame moves. Previous studies demonstrated that the PRRSV sliding sequence was "UUUAAAC", followed by a three necked "pseudoknot" structural sequence that mediates ribosome withdrawal. When the ribosome in the translational extension step moves to the sliding sequence, the tRNA is detached from the ribosome, and after the ribosome is reversed by one step in a sliding manner, the tRNA is caused to re-enter the reading frame "-1Ribosomal Frameshifting", at which time the peptide acyl transfer center is unaffected, detachment of the nascent peptide chain does not occur, and the ribosome enters the translation process of ORF1 b. The phenomenon of minus 1Ribosomal Frameshifting can lead the virus to have the characteristics of simple genome, high replication speed and high utilization rate of genetic materials, and is beneficial to propagation and growth of the virus. If the process of PRRSV-1 Ribosomal Frameshifting can be inhibited, the replication of PRRSV can be blocked, so as to attain the goal of inhibiting virus proliferation. Therefore, research on drugs capable of effectively inhibiting the-1 Ribosomal Frameshifting process of PRRSV is of great importance in developing drugs against PRRSV or for treating blue-porcine reproductive and respiratory syndrome caused by PRRSV.
5- (4-Chlorophenyl) -6-ethylpyrimidine-2,4-diamine (5- (4-Chlorophenyl) -6-ethylpyrimidine-2, 4-diamine), commonly known as pyrimethamine, CAS 58-14-0, molecular formula C 12 H 14 Cl 2 N 4 The molecular weight is 248.71, and the structural formula is as above. 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine is an anti-hepatitis B virus drug, however, hepatitis B virus is a hepadnavirus genus and is two completely different viruses from PRRSV, and no report has been made on the use of 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine for inhibiting PRRSV and treating porcine reproductive and respiratory syndrome.
Disclosure of Invention
The invention aims to provide an application of pyrimidine derivatives shown in a formula I in preparing medicines for resisting porcine reproductive and respiratory syndrome virus.
The invention provides application of a compound shown in a formula I or a salt thereof in preparing a medicament for resisting porcine reproductive and respiratory syndrome virus:
wherein M is N or CH;
R 1 selected from H or NH 2 ；
R 2 Selected from the group consisting ofn is 0, 1, 2 or 3, m is 0, 1, 2 or 3, R 4 Each independently selected from H, halogen or C 1-6 An alkyl group; r is R 3 Selected from H or C 1-6 An alkyl group;
or R is 2 、R 3 Connection formationR 5 Selected from H, halogen or C 1-6 An alkyl group.
Further, the structure of the compound is shown as a formula II:
wherein R is 3 Selected from H or C 1-4 Alkyl, R 4 Each independently selected from H, halogen or C 1-4 An alkyl group. Further, the compound is selected from:
further, the compound is:
further, the medicament is a medicament for preventing and/or treating porcine reproductive and respiratory syndrome.
Further, the drug is a drug for inhibiting the frame shift of the ribosome at the-1 position of the porcine reproductive and respiratory syndrome virus.
Further, the drug is a drug that blocks replication of porcine reproductive and respiratory syndrome virus.
Further, the drug is a drug for inhibiting proliferation of porcine reproductive and respiratory syndrome virus.
The invention discovers for the first time that 5- (4-chlorphenyl) -6-ethylpyrimidine-2,4-diamine has remarkable inhibition and blocking effects on the PRRSV-1 Ribosomal Frameshifting process, can effectively inhibit the replication of PRRSV and inhibit the proliferation of PRRSV, and the 5- (4-chlorphenyl) -6-ethylpyrimidine-2,4-diamine can be used for preparing anti-PRRSV medicaments and medicaments for treating blue pig ear diseases caused by PRRSV, and has wide application prospect.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
FIG. 1.schematic diagram of experimental flow and experimental results of inhibition of PRRSV ribosomal frameshifting process by 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine in a luciferase reporter system. Wherein a is a design scheme of a report system, when the frame shift of a ribosome normally occurs, the report genes Renilla and Firefly are both expressed, and when the frame shift of the ribosome is prevented, the report genes Renilla are expressed, and the Firefly is not expressed; b is a report system workflow, firstly, stably integrating a report vector into a host cell (pig kidney cell PK 15) through a lentiviral vector, carrying out compound treatment on a positive monoclonal cell, and judging the inhibiting effect of the compound on the PRRSV ribosome frameshift process through a double-luciferase report gene detection kit and an enzyme-labeled instrument; c is the inhibiting effect of 10 mu M of 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine on ribosome frameshift in a double luciferase reporter gene system.
FIG. 2. Inhibition effect of different concentrations of 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine on PRRSV ribosomal frameshifting.
FIG. 3 shows the results of cytotoxicity experiments of 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine at different concentrations.
FIG. 4.schematic diagram of experimental flow and experimental results of the inhibition of PRRSV ribosomal frameshifting process by 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine in a fluorescent protein reporter system. Wherein a is a fluorescent protein report system design scheme, when the frame shift of the ribosome normally happens, both the reporter gene GFP and the RFP are expressed, and when the frame shift of the ribosome is prevented, the reporter gene GFP is expressed, and the RFP is not expressed; b is a report system workflow, firstly, stably integrating a report vector into a host cell (pig kidney cell PK 15) through a lentiviral vector, carrying out compound treatment on a positive monoclonal cell, obtaining a fluorescence image through a fluorescence microscope, and judging the inhibition effect of the compound on the PRRSV ribosome frameshift process through fluorescence signal analysis; c is the expression of GFP and RFP after treatment with 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine (fluorescent image); d is the quantitative analysis of the fluorescence signal value of the graph c.
FIG. 5.5 inhibition effect of 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine on PRRSV live virus, wherein NC is a negative control without virus.
Detailed Description
The materials and equipment used in the embodiments of the present invention are all known products and are obtained by purchasing commercially available products.
Example 1, inhibition of PRRSV-1 Ribosomal Frameshifting Process by 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine (luciferase reporter)
1. Experimental method
The inhibition of PRRSV-1 Ribosomal Frameshifting process by drugs was investigated using in vitro cultured cells. Firstly, constructing a double-luciferase reporter gene lentiviral vector containing a PRRSV virus genome sliding sequence, and detecting the action effect of a drug in a pig kidney cell line PK15, wherein the drug concentration is 10 mu M.
1.1 vector construction:
(1) The gene sequence of the sliding region of PRRSV virus-1 Ribosomal Frameshifting is synthesized. Double enzyme digestion is carried out on the lentiviral skeleton vector, and enzyme digestion site selection is carried out: ecoRI+BamHI. Enzyme cutting conditions: 37℃for 15 minutes. And after the enzyme digestion is finished, recovering enzyme digestion products through nucleic acid electrophoresis. For the synthetic PRRSV virus-1 Ribosomal Frameshifting sliding region gene sequence, denaturation at 95℃is first carried out for 10 minutes, annealing at 72℃is carried out for 30 seconds, and then the annealed product is mixed with the cleaved product in the following mixing ratio: cleavage product = 3:1, 10 μl of T4 ligase was added to the above mixture and ligated in a 16 ℃ constant temperature metal bath for 16 hours. Plasmid transformation competent cells: and taking out competent cells from a refrigerator at the temperature of minus 80 ℃, melting on ice, adding 10 microliters of plasmids (the plasmids are slow virus vectors carrying the gene sequences of the PRRSV virus-1 Ribosomal Frameshifting sliding region) into 100 microliters of competent cells, adding the plasmids, uniformly mixing, standing on ice for 30 minutes, putting the mixture of the competent cells and the plasmids into water at the temperature of 42 ℃, and carrying out heat shock for 90 seconds. After the heat shock was completed, the mixture of competent cells and plasmid was cooled on ice for 10 seconds. Transferring the cooled mixed solution to a solid LB plate, uniformly coating the liquid on the surface of a solid LB culture medium by using a glass rod, inverting the solid LB culture medium, and culturing in a incubator at 37 ℃ for 16 hours. After the cultivation, the monoclonal bacterial plaque is picked up by a pipette tip and inoculated with a liquid LB medium, and after the cultivation for 8 hours in a shaking table at 37 ℃, the bacterial plaque is sent to a company for sequencing. And performing amplification culture on the monoclonal with correct sequencing results, and performing plasmid extraction.
(2) Renilla luciferase was constructed upstream of the-1 Ribosomal Frameshifting region and firefly luciferase was constructed downstream of the-1 Ribosomal Frameshifting region by homologous recombination. The specific method comprises the following steps: the Renilla luciferase gene sequence was amplified by PCR and homology arms were added upstream and downstream of the sequence, respectively. And (3) carrying out PCR on the vector obtained in the step (1) to obtain a linearization vector. And obtaining a renilla luciferase gel recovery product and a linearization carrier gel recovery product through nucleic acid electrophoresis and gel recovery. Homologous recombination: the renilla luciferase gel recovery product and the linearized carrier gel recovery product were mixed in a mass ratio of 3:1, and 2. Mu.l of homologous recombinase was added for reaction at 37℃for 15 minutes.
(3) Plasmid transformation competent cells are selected, monoclonal and sequenced, and the virus skeleton vector with correct sequencing is amplified and plasmid extraction is carried out. The competent cells were transformed and the extraction procedure was the same as in step (1).
1.2 establishing a screening cell system:
(1) The viral backbone vector containing the double luciferase and the-1 Ribosomal Frameshifting region, PMD2.G and plpax2 packaging vector are mixed with PEI according to the volume ratio of 4:2:1, 70 microgram PEI is added to 35 microgram DNA, and 293T cells are transfected.
(2) The supernatant of 293T cell culture after 48h and 72h transfection of viral plasmid was collected, centrifuged at 12000g for 10min, and after removal of cellular impurities, lentiviral particles were collected by cesium chloride gradient centrifugation.
(3) Lentiviral particles were added to PK15 cells and, 7 days after infection, puro screened positive cells. The cells were digested with pancreatin to prepare single cell suspensions, and monoclonal sorting was performed by flow cytometry.
(4) Genotyping the cultured monoclonal cells and performing expansion culture on the positive clones to obtain PK15 cells (CMV-Renilla-framshift-Firefly) containing the-1 Ribosomal Frameshifting region.
1.3 experimental procedure for drug inhibition-1 Ribosomal Frameshifting procedure:
(1) PK15 cells containing the-1 Ribosomal Frameshifting region were cultured in 96-well plates, and the drugs to be tested were added thereto at a drug concentration of 10. Mu.M, respectively.
(2) After 8h of culture, the cells were lysed, and firefly luciferase substrate was added to the lysate, and the luminescence value was detected by an ELISA reader.
(3) Adding a renilla luciferase substrate, and detecting a luminescence value in an enzyme-labeled instrument.
(4) The ratio of firefly luciferase to Renilla luciferase was used as a reference for the efficiency of-1 Ribosomal Frameshifting. A larger ratio indicates a higher efficiency of-1 Ribosomal Frameshifting, a lower ratio indicates a lower efficiency of-1 Ribosomal Frameshifting, i.e., a lower ratio indicates a higher efficiency of the drug to inhibit the-1 Ribosomal Frameshifting process, which can better inhibit the-1 Ribosomal Frameshifting process.
2. Experimental results
FIG. 1c shows the inhibitory effect of 10. Mu.M 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine on PRRSV in the course of-1 Ribosomal Frameshifting. To further determine the effect of different concentrations of 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine on PRRSV virus-1 Ribosomal Frameshifting efficiency, the efficacy of PRRSV virus-1 Ribosomal Frameshifting at different 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine concentrations was studied using the luciferase-based-1 Ribosomal Frameshifting efficacy screening system and method described previously, and the relationship between 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine concentration changes and the inhibitory effect of-1 Ribosomal Frameshifting was analyzed by firefly luciferase activity. The detection result shows that the 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine can obviously inhibit the efficiency of-1 Ribosomal Frameshifting at the concentration of 1 mu M (figure 2).
To further determine the inhibition of PRRSV virus-1 Ribosomal Frameshifting by 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine, cytotoxicity assays were performed in wild type MARC-145 cells. The results showed that 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine was not toxic to wild-type MARC-145 cells at concentrations of 0.25. Mu.M, 0.5. Mu.M, 1. Mu.M, 2.5. Mu.M, 5. Mu.M, and 10. Mu.M in MARC-145 cells (FIG. 3).
The experimental results show that: 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine has an inhibitory effect on the-1 Ribosomal Frameshifting process of PRRSV virus, at a minimum effective concentration of 1. Mu.M, when the compound is not toxic to wild type MARC-145 cells.
Example 2, inhibition of PRRSV-1 Ribosomal Frameshifting Process by 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine (fluorescent protein reporter System)
1. Experimental method
To further determine the inhibitory activity of 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine on PRRSV-1 Ribosomal Frameshifting process, the present invention constructs a reporter gene system based on short half-life fluorescent proteins. Firstly, ubiquitin is connected to RFP and GFP to obtain UbRFP and UbGFP, firefly luciferase and Renilla luciferase in a dual luciferase report system are replaced by Ub-RFP and Ub-GFP respectively, and the updated report system is as follows: CMV-UbRFP-Frameshift-UbGFP. In detection, CMV-UbRFP-Frameshift-UbGFP cells are passaged in a 96-well plate 24h in advance, so that the cell fusion degree is preferably 80% -90% during dosing. 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine (2. Mu.M) was added to the petri dish for 1 hour, after which 5. Mu.MMG-132 (proteasome inhibitor) was added for 3 hours and 3 duplicate wells were placed in each group. Finally, fluorescence ratio information of GFP and RFP is obtained through a fluorescence microscope.
2. Experimental results
The results showed that 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine still had a significant inhibitory effect on PRRSV-1 Ribosomal Frameshifting process in the fluorescent protein reporter system (fig. 4).
Example 3, inhibitory Activity of 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine against live viruses of PRRSV
1. Experimental method
In order to further determine the inhibition activity of 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine on the in-vivo PRRSV, the invention constructs an in-vitro culture and detection system of PRRSV. First, MARC145 cells were grown at 2X 10 in biosafety class II laboratory 5 Each ml was inoculated into a 6-well plate, and after culturing for 24 hours, cells were infected with 0.1MOI virus per well, and after 24 hours, 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine (10. Mu.M) or DMSO was added. No virus was added to the negative control group (NC group). After culturing in a cell culture box at 37 ℃ for 48 hours, collecting cells, extracting total RNA of the cells by a Trizol method, obtaining cDNA by a reverse transcription kit, and detecting PRRSV virus copy number by a qPCR method. qPCR primers were as follows: PRRSVORF7-FAAACCAGTCCAGAGGCAAGG (SEQ ID NO: 1), PRRSVORF7-RGCAAACTAAACTCCACAGTGTAA (SEQ ID NO: 2); GAPDH-FGAAGGTGAAGGTCGGAGTCA (SEQ ID NO: 3), GAPDH-RCATGTAAACCATGTAGTTGAGGTC (SEQ ID NO: 4).
2. Experimental results
The results showed that 10. Mu.M of 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine had a significant inhibitory effect on PRRSV virus proliferation in vitro culture (FIG. 5).
In summary, the invention provides application of pyrimidine derivatives in preparing medicines for resisting porcine reproductive and respiratory syndrome virus. The compound has remarkable inhibiting and blocking effects on the-1-position ribosome frameshift process of the porcine reproductive and respiratory syndrome virus, can effectively inhibit the replication of PRRSV, inhibits the proliferation of PRRSV, and the 5- (4-chlorophenyl) -6-ethylpyrimidine-2,4-diamine can be used for preparing a medicament for resisting PRRSV and a medicament for treating blue pig ear disease caused by PRRSV, and has wide application prospect.
Claims (8)
1. The use of a compound of formula I or a salt thereof for the manufacture of a medicament against porcine reproductive and respiratory syndrome virus:
wherein M is N or CH;
R 1 selected from H or NH 2 ；
R 2 Selected from the group consisting ofn is 0, 1, 2 or 3, m is 0, 1, 2 or 3, R 4 Each independently selected from H, halogen or C 1-6 An alkyl group; r is R 3 Selected from H or C 1-6 An alkyl group;
or R is 2 、R 3 Connection formationR 5 Selected from H, halogen or C 1-6 An alkyl group.
2. The use according to claim 1, wherein the compound has the structure according to formula II:
wherein R is 3 Selected from H or C 1-4 Alkyl, R 4 Each independently selected from H, halogen or C 1-4 An alkyl group.
3. Use according to claim 1, characterized in that said compound is selected from:
4. use according to claim 3, characterized in that the compound is:
5. the use according to any one of claims 1 to 4, wherein the medicament is a medicament for the prevention and/or treatment of porcine reproductive and respiratory syndrome.
6. The use according to any one of claims 1 to 4, wherein the medicament is a medicament for inhibiting the frame shift of the ribosome-1 in porcine reproductive and respiratory syndrome virus.
7. The use according to any one of claims 1 to 4, wherein the medicament is a medicament for blocking replication of porcine reproductive and respiratory syndrome virus.
8. The use according to any one of claims 1 to 4, wherein the medicament is a medicament for inhibiting proliferation of porcine reproductive and respiratory syndrome virus.
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